U.S. patent application number 13/958424 was filed with the patent office on 2013-12-05 for inhibitors of flaviviridae viruses.
This patent application is currently assigned to Gilead Sciences, Inc.. The applicant listed for this patent is Gilead Sciences, Inc.. Invention is credited to Michael Mertzman Belmont, Eda Canales, Lee S. Chong, Michael O'Neil Hanrahan Clarke, Edward Doerffler, Scott E. Lazerwith, Willard Lew, Philip Anthony Morganelli, William J. Watkins.
Application Number | 20130323203 13/958424 |
Document ID | / |
Family ID | 43827460 |
Filed Date | 2013-12-05 |
United States Patent
Application |
20130323203 |
Kind Code |
A1 |
Canales; Eda ; et
al. |
December 5, 2013 |
INHIBITORS OF FLAVIVIRIDAE VIRUSES
Abstract
Provided are compounds of Formula I: ##STR00001## and
pharmaceutically acceptable salts and esters thereof. The
compounds, compositions, and methods provided are useful for the
treatment of Flaviviridae virus infections, particularly hepatitis
C infections.
Inventors: |
Canales; Eda; (San Mateo,
CA) ; Chong; Lee S.; (Newark, CA) ; Clarke;
Michael O'Neil Hanrahan; (Redwiid City, CA) ;
Doerffler; Edward; (Union City, CA) ; Lazerwith;
Scott E.; (San Francisco, CA) ; Lew; Willard;
(San Mateo, CA) ; Belmont; Michael Mertzman;
(Belmont, CA) ; Morganelli; Philip Anthony;
(Oakland, CA) ; Watkins; William J.; (Saratoga,
CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Gilead Sciences, Inc. |
Foster City |
CA |
US |
|
|
Assignee: |
Gilead Sciences, Inc.
Foster City
CA
|
Family ID: |
43827460 |
Appl. No.: |
13/958424 |
Filed: |
August 2, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13006761 |
Jan 14, 2011 |
8524764 |
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13958424 |
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61295576 |
Jan 15, 2010 |
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Current U.S.
Class: |
424/85.4 ;
514/210.18; 514/252.01; 514/255.05; 514/316; 514/321; 514/326;
514/370; 514/422; 514/43; 514/444; 514/448; 544/238; 544/405;
546/187; 546/197; 546/208; 548/194; 548/518; 548/526; 549/60;
549/71 |
Current CPC
Class: |
A61P 31/00 20180101;
A61K 31/4025 20130101; A61K 31/381 20130101; A61K 31/497 20130101;
A61K 45/06 20130101; A61K 31/4545 20130101; C07D 409/14 20130101;
A61P 31/12 20180101; A61K 31/501 20130101; C07D 333/38 20130101;
A61K 31/427 20130101; A61K 31/4535 20130101; A61P 31/14 20180101;
A61K 31/454 20130101; C07D 493/04 20130101; C07D 413/12 20130101;
C07D 409/12 20130101 |
Class at
Publication: |
424/85.4 ;
546/187; 514/316; 514/43; 549/60; 514/444; 548/518; 514/422;
548/526; 546/197; 514/321; 546/208; 514/326; 514/210.18; 544/405;
514/255.05; 548/194; 514/370; 544/238; 514/252.01; 549/71;
514/448 |
International
Class: |
C07D 333/38 20060101
C07D333/38; A61K 31/4545 20060101 A61K031/4545; A61K 45/06 20060101
A61K045/06; C07D 409/12 20060101 C07D409/12; A61K 31/381 20060101
A61K031/381; A61K 31/501 20060101 A61K031/501; C07D 493/04 20060101
C07D493/04; A61K 31/4535 20060101 A61K031/4535; A61K 31/454
20060101 A61K031/454; A61K 31/497 20060101 A61K031/497; C07D 413/12
20060101 C07D413/12; A61K 31/427 20060101 A61K031/427; C07D 409/14
20060101 C07D409/14; A61K 31/4025 20060101 A61K031/4025 |
Claims
1.-21. (canceled)
22. A compound of Formula I: ##STR00157## or a pharmaceutically
acceptable salt or ester thereof, wherein: R.sup.1 is C.sub.1-12
alkyl; each Q.sup.1 is independently selected from the group
consisting of C.sub.1-6 alkyl, 5-10 membered heteroaryloxy, 4-12
membered heterocyclyloxy, --OH, and --C(O)OR.sup.10, wherein said
heteroaryl portion of said 5-10 membered heteroaryloxy and said
heterocyclyl portion of said 4-12 membered heterocyclyloxy each
comprises one to four heteroatoms selected from O, S, or N;
R.sup.10 is selected from the group consisting of H, and C.sub.1-12
alkyl; R.sup.2 is optionally substituted C.sub.3-12 cycloalkyl;
wherein, each substituted R.sup.2 is substituted with one or more
C.sub.1-6 alkyl; each Q.sup.2, independently, is selected from the
group consisting of --S(O).sub.2R.sup.20, optionally substituted
C.sub.1-6 alkyl, OH, --C(O)OR.sup.20, and --CN; each R.sup.20,
independently, is selected from the group consisting of H, and
C.sub.1-12 alkyl; Y is --R.sup.3-L-Het, --N(R.sup.4)(R.sup.5) or
--R.sup.6.dbd.NOR.sup.7; R.sup.3 is selected from the group
consisting of C.sub.1-12 alkylene, C.sub.3-12 cycloalkylene, and
3-12 membered heterocyclylene, wherein said 3-12 membered
heterocyclylene comprises one to four heteroatoms selected from O,
S, or N; L is selected from the group consisting of
--OC(O)N(R.sup.4)--, --N(R.sup.4)C(O)O--, --N(R.sup.4)S(O).sub.2--,
--N(R.sup.4)C(O)--, --C(O)--, --C(O)O--, --OC(O)--,
--N(R.sup.4)N(R.sup.4)C(O)O--, and --N(R.sup.4)N(R.sup.4)--; Het is
an optionally substituted 3-12 membered heterocyclyl or optionally
substituted 5-10 membered heteroaryl, wherein said optionally
substituted 3-12 membered heterocyclyl or optionally substituted
5-10 membered heteroaryl comprises one to four heteroatoms selected
from O, S, or N; wherein, each substituted Het is substituted with
one or more Q.sup.4; each Q.sup.4, independently, is selected from
the group consisting of halogen, oxo, C.sub.1-6 alkyl, and
--C(O)OR.sup.40; R.sup.40 is C.sub.1-12 alkyl; each R.sup.4 is
independently selected from the group consisting of H,
C.sub.1-C.sub.12 alkyl, C.sub.3-C.sub.12 cycloalkyl, 5-10 membered
heteroaryl and 4-10 membered heterocyclyl wherein each
C.sub.1-C.sub.12 alkyl, C.sub.3-C.sub.12 cycloalkyl, -5-10 membered
heteroaryl or 4-10 membered heterocyclyl is optionally substituted
with one or more Q.sup.1, wherein said 5-10 membered heteroaryl and
said 4-10 membered heterocyclyl each comprises one to four
heteroatoms selected from O, S, or N; each R.sup.5 is independently
selected from the group consisting of C.sub.1-C.sub.12 alkyl,
C.sub.3-C.sub.12 cycloalkyl, 5-10 membered heteroaryl and 4-10
membered heterocyclyl wherein each C.sub.1-C.sub.12 alkyl,
C.sub.3-C.sub.12 cycloalkyl, 5-10 membered heteroaryl or 4-10
membered heterocyclyl is optionally substituted with one or more
Q.sup.1, wherein said 5-10 membered heteroaryl and said 4-10
membered heterocyclyl each comprises one to four heteroatoms
selected from O, S, or N; R.sup.6 is C.sub.3-C.sub.12
cycloalkylyne; and R.sup.7 is selected from the group consisting of
optionally substituted C.sub.1-12 alkyl, optionally substituted
C.sub.6-14 aryl, optionally substituted 6-11 membered
heteroarylalkyl, and optionally substituted C.sub.7-11 arylalkyl,
wherein said heteroaryl portion of said 6-11 membered
heteroarylalkyl comprises one to four heteroatoms selected from O,
S, or N; wherein, each substituted R.sup.7 is substituted with one
or more Q.sup.2.
23. The compound of claim 22, wherein R.sup.1 is optionally
substituted C.sub.3-C.sub.7 secondary or tertiary alkyl.
24. The compound of claim 22, wherein R.sup.2 is optionally
substituted methylcyclohexyl or optionally substituted
methylcyclohexenyl.
25. The compound of claim 22 represented by Formula II:
##STR00158## or pharmaceutically acceptable salts or esters
thereof, wherein R.sup.2 is optionally substituted
4-methylcyclohexyl or optionally substituted
4-methylcyclohexenyl.
26. The compound of claim 22 wherein R.sup.2 is: ##STR00159##
27. The compound of claim 22 wherein R.sup.2 is: ##STR00160##
28. The compound of claim 22, wherein Y is --R.sup.3-L-Het.
29. The compound of claim 28, wherein R.sup.3 is selected from the
group consisting of optionally substituted C.sub.1-6 alkylene,
C.sub.4-6 cycloalkylene, and 5-6 membered heterocyclylene.
30. The compound of claim 28 wherein Het is selected from the group
consisting of optionally substituted pyridinyl, optionally
substituted pyridazinyl, optionally substituted
tetrahydro-2H-pyranyl, optionally substituted piperidinyl,
optionally substituted pyrrolidinyl, optionally substituted
tetrahydrothiophenyl, optionally substituted pyrazinyl, optionally
substituted 1H-tetrazolyl, optionally substituted azetidinyl,
optionally substituted tetrahydrofuranyl, optionally substituted
tetrahydro-2H-furo[2,3-b]furanyl, optionally substituted thiazoyl,
optionally substituted 1H-imidazolyl, optionally substituted
4H-1,2,4-triazolyl, optionally substituted 1H-pyrazolyl, optionally
substituted 1,3,4-thiadiazolyl, optionally substituted quinolinyl,
optionally substituted [1,2,4]triazolo[4,3-a]pyridinyl, optionally
substituted thiophenyl, optionally substituted 1,2,4-thiadiazolyl,
optionally substituted pyrimidinyl, optionally substituted
1H-1,2,3-triazolyl, optionally substituted 1,3,4-oxadiazolyl, and
optionally substituted imidazo[1,2-b]pyridazinyl.
31. The compound of claim 22 wherein Y is
--N(R.sup.4)(R.sup.5).
32. The compound of claim 31 wherein R.sup.4 is selected from the
group consisting of H and optionally substituted C.sub.1-C.sub.6
alkyl.
33. The compound of claim 31 wherein R.sup.5 is selected from the
group consisting of C.sub.1-C.sub.6 alkyl, C.sub.5-C.sub.6
cycloalkyl, 5-10 membered heteroaryl and 4-10 membered heterocyclyl
wherein each C.sub.1-C.sub.6 alkyl, C.sub.5-C.sub.6 cycloalkyl,
5-10 membered heteroaryl or 4-10 membered heterocyclyl is
optionally substituted with one or more Q.sup.1.
34. The compound of claim 22 wherein Y is
--R.sup.6.dbd.NOR.sup.7.
35. The compound of claim 34 wherein R.sup.6 is cyclohexylyne.
Description
[0001] This application is filed under 35 U.S.C. 111(a) claiming
the benefit under 35 U.S.C. 119(e) of U.S. provisional application
61/295,576 filed Jan. 15, 2010 which is herein incorporated by
reference in its entirety for all purposes.
FIELD OF THE INVENTION
[0002] The present application includes novel inhibitors of
Flaviviridae viruses, compositions containing such compounds,
therapeutic methods that include the administration of such
compounds.
BACKGROUND OF THE INVENTION
[0003] Viruses comprising the Flaviviridae family include at least
three distinguishable genera including pestiviruses, flaviviruses,
and hepaciviruses (Calisher, et al., J. Gen. Virol., 1993, 70,
37-43). While pestiviruses cause many economically important animal
diseases such as bovine viral diarrhea virus (BVDV), classical
swine fever virus (CSFV, hog cholera) and border disease of sheep
(BDV), their importance in human disease is less well characterized
(Moennig, V., et al., Adv. Vir. Res. 1992, 48, 53-98). Flaviviruses
are responsible for important human diseases such as dengue fever
and yellow fever while hepaciviruses cause hepatitis C virus
infections in humans. Other important viral infections caused by
the Flaviviridae family include West Nile virus (WNV) Japanese
encephalitis virus (JEV), tick-borne encephalitis virus, Junjin
virus, Murray Valley encephalitis, St Louis enchaplitis, Omsk
hemorrhagic fever virus and Zika virus.
[0004] The hepatitis C virus (HCV) is the leading cause of chronic
liver disease worldwide (Boyer, N. et al. J. Hepatol. 32:98-112,
2000) so a significant focus of current antiviral research is
directed toward the development of improved methods of treatment of
chronic HCV infections in humans (Di Besceglie, A. M. and Bacon, B.
R., Scientific American, October: 80-85, (1999); Gordon, C. P., et
al., J. Med. Chem. 2005, 48, 1-20; Maradpour, D.; et al., Nat. Rev.
Micro. 2007, 5(6), 453-463). A number of HCV treatments are
reviewed by Bymock et al. in Antiviral Chemistry &
Chemotherapy, 11:2; 79-95 (2000). Virologic cures of patients with
chronic HCV infection are difficult to achieve because of the
prodigious amount of daily virus production in chronically infected
patients and the high spontaneous mutability of HCV virus (Neumann,
et al., Science 1998, 282, 103-7; Fukimoto, et al., Hepatology,
1996, 24, 1351-4; Domingo, et al., Gene, 1985, 40, 1-8; Martell, et
al., J. Virol. 1992, 66, 3225-9.
[0005] Currently, there are primarily two antiviral compounds,
ribavirin, a nucleoside analog, and interferon-alpha (.alpha.)
(IFN), that are used for the treatment of chronic HCV infections in
humans. Ribavirin alone is not effective in reducing viral RNA
levels, has significant toxicity, and is known to induce anemia.
The combination of IFN and ribavirin has been reported to be
effective in the management of chronic hepatitis C (Scott, L. J.,
et al. Drugs 2002, 62, 507-556) but less than half the patients
given this treatment show a persistent benefit.
[0006] Combined, infections from the Flaviviridae virus family
cause significant mortality, morbidity and economic losses
throughout the world. Alkynyl substituted thiophenes with
anti-Flaviviridae virus activity have been disclosed by Chan, et
al., WO 2008058393; Wunberg, et al., WO 2006072347; and Chan, et
al., WO 2002100851; but none of these are currently clinically
approved antiviral therapeutics. Therefore, there remains a need to
develop effective treatments for Flaviviridae virus infections.
SUMMARY OF THE INVENTION
[0007] Provided are compounds of Formula I:
##STR00002##
or a pharmaceutically acceptable salt or ester thereof,
wherein:
[0008] R.sup.1 is selected from the group consisting of optionally
substituted C.sub.1-12 alkyl, optionally substituted C.sub.2-12
alkenyl, optionally substituted C.sub.2-12 alkynyl, optionally
substituted C.sub.3-12 cycloalkyl, optionally substituted
C.sub.6-14 aryl, optionally substituted 3-14 membered heteroaryl,
optionally substituted 3-12 membered heterocyclyl, optionally
substituted 3-18 membered heteroarylalkyl, optionally substituted
3-18 membered heterocyclylalkyl and optionally substituted
C.sub.6-18 arylalkyl, wherein, each substituted R.sup.1 is
substituted with one or more Q.sup.1;
[0009] each Q.sup.1 is independently selected from the group
consisting of halogen, oxo, oxide, --NO.sub.2, --N(.dbd.O),
--SR.sup.10, --S(O)R.sup.10, --S(O).sub.2R.sup.10,
--S(O).sub.2NR.sup.10R.sup.11, --NR.sup.10C(O)R.sup.11,
--NR.sup.10C(O)NR.sup.11R.sup.12, --NR.sup.10S(O)R.sup.11,
--NR.sup.10S(O).sub.2R.sup.11, --OP(O)R.sup.11R.sup.12,
--P(O)R.sup.11R.sup.12, --P(O)OR.sup.11R.sup.12,
--P(O)(OR.sup.11)OR.sup.12, --C(O)NR.sup.11R.sup.12, optionally
substituted C.sub.1-6 alkyl, optionally substituted C.sub.2-6
alkenyl, optionally substituted C.sub.2-6 alkynyl, optionally
substituted C.sub.3-6 cycloalkyl, optionally substituted C.sub.6-12
arylalkyl, optionally substituted C.sub.6-12 aryl, optionally
substituted 3-14 membered heteroaryl, optionally substituted
C.sub.1-6 alkyloxy, optionally substituted C.sub.2-6 alkenyloxy,
optionally substituted C.sub.2-6 alkynyloxy, optionally substituted
C.sub.3-6 cycloalkyloxy, optionally substituted C.sub.6-12 aryloxy,
optionally substituted 3-14 membered heteroaryloxy, optionally
substituted 4-12 membered heterocyclyloxy, optionally substituted
--C(O)C.sub.1-6 alkyl, optionally substituted --C(O)C.sub.2-6
alkenyl, optionally substituted --C(O)C.sub.2-6 alkynyl, optionally
substituted --C(O)C.sub.3-6 cycloalkyl, optionally substituted
--C(O)C.sub.6-12 aryl, optionally substituted --C(O)-3-14 membered
heteroaryl, optionally substituted --C(O)C.sub.6-12 arylalkyl,
optionally substituted-3-10 membered heterocyclyl, --OH,
--NR.sup.11R.sup.12, --C(O)OR.sup.10, --CN, --N.sub.3,
--C(.dbd.NR.sup.13)NR.sup.11R.sup.12, --C(.dbd.NR.sup.13)OR.sup.10,
--NR.sup.10C(.dbd.NR.sup.13)NR.sup.11R.sup.12,
--NR.sup.11C(O)OR.sup.10, and --OC(O)NR.sup.11R.sup.12;
[0010] each R.sup.10, R.sup.11, and R.sup.12, independently, is
selected from the group consisting of H, optionally substituted
C.sub.1-12 alkyl, optionally substituted C.sub.2-12 alkenyl,
optionally substituted C.sub.2-12 alkynyl, optionally substituted
C.sub.3-12 cycloalkyl, optionally substituted C.sub.6-14 aryl,
optionally substituted 3-14 membered heteroaryl, optionally
substituted 3-12 membered heterocyclyl, optionally substituted 3-18
membered heteroarylalkyl, and optionally substituted C.sub.6-18
arylalkyl;
[0011] or R.sup.11 and R.sup.12 taken together with the atoms to
which they are attached form a 3 to 10 membered heterocyclyl;
[0012] each R.sup.13, independently, is selected from the group
consisting of H, optionally substituted C.sub.1-12 alkyl,
optionally substituted C.sub.2-12 alkenyl, optionally substituted
C.sub.2-12 alkynyl, optionally substituted C.sub.3-12 cycloalkyl,
optionally substituted C.sub.6-14 aryl, optionally substituted 3-14
membered heteroaryl, optionally substituted 3-12 membered
heterocyclyl, optionally substituted 3-18 membered heteroarylalkyl,
optionally substituted C.sub.6-18 arylalkyl, --CN, --C(O)R.sup.14,
--CHO and --S(O).sub.2R.sup.14;
[0013] each R.sup.14, independently, is optionally substituted
C.sub.1-12 alkyl;
[0014] wherein, each substituted Q.sup.1, substituted R.sup.10,
substituted R.sup.11, substituted R.sup.12, substituted R.sup.13,
or substituted R.sup.14 is independently substituted with one or
more Q.sup.6;
[0015] R.sup.2 is selected from the group consisting of optionally
substituted C.sub.1-12 alkyl, optionally substituted C.sub.2-12
alkenyl, optionally substituted C.sub.2-12 alkynyl, optionally
substituted C.sub.3-12 cycloalkyl, optionally substituted
C.sub.6-14 aryl, optionally substituted 3-14 membered heteroaryl,
optionally substituted 3-12 membered heterocyclyl, optionally
substituted 3-18 membered heteroarylalkyl, and optionally
substituted C.sub.6-18 arylalkyl;
[0016] wherein, each substituted R.sup.2 is substituted with one or
more Q.sup.2;
[0017] each Q.sup.2, independently, is selected from the group
consisting of halogen, oxo, oxide, --NO.sub.2, --N(.dbd.O),
--SR.sup.20, --S(O)R.sup.20, --S(O).sub.2R.sup.20,
--S(O).sub.2NR.sup.20R.sup.21, --NR.sup.20C(O)R.sup.21,
--NR.sup.20C(O)NR.sup.21R.sup.22, --NR.sup.20S(O)R.sup.21,
--NR.sup.20S(O).sub.2R.sup.21, --OP(O)R.sup.21R.sup.22,
--P(O)R.sup.21R.sup.22, --P(O)OR.sup.21R.sup.22,
--P(O)(OR.sup.21)OR.sup.22, --C(O)NR.sup.21R.sup.22, optionally
substituted C.sub.1-6 alkyl, optionally substituted C.sub.2-6
alkenyl, optionally substituted C.sub.2-6 alkynyl, optionally
substituted C.sub.3-6 cycloalkyl, optionally substituted C.sub.6-12
arylalkyl, optionally substituted C.sub.6-12 aryl, optionally
substituted 3-14 membered heteroaryl, optionally substituted
C.sub.1-6 alkyloxy, optionally substituted C.sub.2-6 alkenyloxy,
optionally substituted C.sub.2-6 alkynyloxy, optionally substituted
C.sub.3-6 cycloalkyloxy, optionally substituted C.sub.6-12 aryloxy,
optionally substituted 3-14 membered heteroaryloxy, optionally
substituted 4-12 membered heterocyclyloxy, optionally substituted
--C(O)C.sub.1-6 alkyl, optionally substituted --C(O)C.sub.2-6
alkenyl, optionally substituted --C(O)C.sub.2-6 alkynyl, optionally
substituted --C(O)C.sub.3-6 cycloalkyl, optionally substituted
--C(O)C.sub.6-12 aryl, optionally substituted --C(O)-3-14 membered
heteroaryl, optionally substituted --C(O)C.sub.6-12 arylalkyl,
optionally substituted 3-10 membered) heterocyclyl, --OH,
--NR.sup.21R.sup.22, --C(O)OR.sup.20, --CN, --N.sub.3,
--C(.dbd.NR.sup.23)NR.sup.21R.sup.22, --C(.dbd.NR.sup.23)OR.sup.20,
--NR.sup.20C(.dbd.NR.sup.23)NR.sup.21R.sup.22,
--NR.sup.21C(O)OR.sup.20, and --OC(O)NR.sup.21R.sup.22;
[0018] each R.sup.20, R.sup.21, and R.sup.22, independently, is
selected from the group consisting of H, optionally substituted
C.sub.1-12 alkyl, optionally substituted C.sub.2-12 alkenyl,
optionally substituted C.sub.2-12 alkynyl, optionally substituted
C.sub.3-12 cycloalkyl, optionally substituted C.sub.6-14 aryl,
optionally substituted 3-14 membered heteroaryl, optionally
substituted 3-12 membered heterocyclyl, optionally substituted 3-18
membered heteroarylalkyl, and optionally substituted C.sub.6-18
arylalkyl;
[0019] or R.sup.21 and R.sup.22 taken together with the atoms to
which they are attached form a 3 to 10 membered heterocyclyl;
[0020] each R.sup.23, independently, is selected from the group
consisting of H, optionally substituted C.sub.1-12 alkyl,
optionally substituted C.sub.2-12 alkenyl, optionally substituted
C.sub.2-12 alkynyl, optionally substituted C.sub.3-12 cycloalkyl,
optionally substituted C.sub.6-14 aryl, optionally substituted 3-14
membered heteroaryl, optionally substituted 3-12 membered
heterocyclyl, optionally substituted 3-18 membered heteroarylalkyl,
optionally substituted C.sub.6-18 arylalkyl, --CN, --C(O)R.sup.24,
--CHO and --S(O).sub.2R.sup.24;
[0021] each R.sup.24 individually is optionally substituted
C.sub.1-12 alkyl;
[0022] wherein, each substituted Q.sup.2, substituted R.sup.20,
substituted R.sup.21, substituted R.sup.22, substituted R.sup.23,
or substituted R.sup.24 is independently substituted with one or
more Q.sup.6;
[0023] Y is --R.sup.3-L-Het, --N(R.sup.4)(R.sup.5) or
--R.sup.6.dbd.NOR.sup.7;
[0024] R.sup.3 is selected from the group consisting of optionally
substituted C.sub.1-12 alkylene, C.sub.2-12 alkenylene, substituted
C.sub.2-12 alkenylene, C.sub.2-12 alkynylene, substituted
C.sub.2-12 alkynylene, C.sub.3-12 cycloalkylene, substituted
C.sub.3-12 cycloalkylene, C.sub.3-12 cycloalkylalkylene,
substituted C.sub.3-12 cycloalkylalkylene, optionally substituted
C.sub.6-14 arylene, optionally substituted 3-14 membered
heteroarylene, optionally substituted 3-12 membered
heterocyclylene, optionally substituted 3-18 membered
heteroarylalkylene, and optionally substituted C.sub.6-18
arylalkylene;
[0025] wherein each substituted R.sup.3 is substituted with one or
more Q.sup.3;
[0026] each Q.sup.3, independently, is selected from the group
consisting of halogen, oxo, oxide, --NO.sub.2, --N(.dbd.O),
--SR.sup.30, --S(O)R.sup.30, --S(O).sub.2R.sup.30,
--S(O).sub.2NR.sup.30R.sup.31, --NR.sup.30C(O)R.sup.31,
--NR.sup.30C(O)NR.sup.31R.sup.32, --NR.sup.30S(O)R.sup.31,
--NR.sup.30S(O).sub.2R.sup.31, --OP(O)R.sup.31R.sup.32,
--P(O)R.sup.31R.sup.32, --P(O)OR.sup.31R.sup.32,
--P(O)(OR.sup.31)OR.sup.32, --C(O)NR.sup.31R.sup.32, optionally
substituted C.sub.1-6 alkyl, optionally substituted C.sub.2-6
alkenyl, optionally substituted C.sub.2-6 alkynyl, optionally
substituted C.sub.3-6 cycloalkyl, optionally substituted C.sub.6-12
arylalkyl, optionally substituted C.sub.6-12 aryl, optionally
substituted 3-14 membered heteroaryl, optionally substituted
C.sub.1-6 alkyloxy, optionally substituted C.sub.2-6 alkenyloxy,
optionally substituted C.sub.2-6 alkynyloxy, optionally substituted
C.sub.3-6 cycloalkyloxy, optionally substituted C.sub.6-12 aryloxy,
optionally substituted 3-14 membered heteroaryloxy, optionally
substituted 4-12 membered heterocyclyloxy, optionally substituted
--C(O)C.sub.1-6 alkyl, optionally substituted --C(O)C.sub.2-6
alkenyl, optionally substituted --C(O)C.sub.2-6 alkynyl, optionally
substituted --C(O)C.sub.3-6 cycloalkyl, optionally substituted
--C(O)C.sub.6-12 aryl, optionally substituted --C(O)-3-14 membered
heteroaryl, optionally substituted --C(O)C.sub.6-12 arylalkyl,
optionally substituted 3-10 membered heterocyclyl, --OH,
--NR.sup.31R.sup.32, --C(O)OR.sup.30, --CN, --N.sub.3,
--C(.dbd.NR.sup.33)NR.sup.31R.sup.32, --C(.dbd.NR.sup.33)OR.sup.30,
--NR.sup.30C(.dbd.NR.sup.33)NR.sup.31R.sup.32,
--NR.sup.31C(O)OR.sup.30, and --OC(O)NR.sup.31R.sup.32;
[0027] each R.sup.30, R.sup.31, and R.sup.32, independently is
selected from the group consisting of H, optionally substituted
C.sub.1-12 alkyl, optionally substituted C.sub.2-12 alkenyl,
optionally substituted C.sub.2-12 alkynyl, optionally substituted
C.sub.3-12 cycloalkyl, optionally substituted C.sub.6-14 aryl,
optionally substituted 3-14 membered heteroaryl, optionally
substituted 3-12 membered heterocyclyl, optionally substituted 3-18
membered heteroarylalkyl, and optionally substituted C.sub.6-18
arylalkyl;
[0028] or R.sup.31 and R.sup.32 taken together with the atoms to
which they are attached form a 3 to 10 membered heterocyclyl;
[0029] each R.sup.33 independently is selected from the group
consisting of H, optionally substituted C.sub.1-12 alkyl,
optionally substituted C.sub.2-12 alkenyl, optionally substituted
C.sub.2-12 alkynyl, optionally substituted C.sub.3-12 cycloalkyl,
optionally substituted C.sub.6-14 aryl, optionally substituted 3-14
membered heteroaryl, optionally substituted 3-12 membered
heterocyclyl, optionally substituted 3-18 membered heteroarylalkyl,
optionally substituted C.sub.6-18 arylalkyl, --CN, --C(O)R.sup.34,
--CHO and --S(O).sub.2R.sup.34;
[0030] each R.sup.34 individually is optionally substituted
C.sub.1-12 alkyl;
[0031] wherein, each substituted Q.sup.3, substituted R.sup.30,
substituted R.sup.31, substituted R.sup.32, substituted R.sup.33,
or substituted R.sup.34 is independently substituted with one or
more Q.sup.6;
[0032] L is selected from the group consisting of
--OC(O)N(R.sup.4)--, --N(R.sup.4)C(O)O--, --N(R.sup.4)S(O).sub.2--,
--N(R.sup.4)C(O)--, --C(O)--, --C(O)O--, --OC(O)--,
--N(R.sup.4)N(R.sup.4)C(O)O--, and --N(R.sup.4)N(R.sup.4)--;
[0033] Het is an optionally substituted 3-12 membered heterocyclyl
or optionally substituted 3-14 membered heteroaryl;
[0034] wherein, each substituted Het is substituted with one or
more Q.sup.4;
[0035] each Q.sup.4, independently, is selected from the group
consisting of halogen, oxo, oxide, --NO.sub.2, --N(.dbd.O),
--SR.sup.40, --S(O)R.sup.40, --S(O).sub.2R.sup.40,
--S(O).sub.2NR.sup.40R.sup.41, --NR.sup.40C(O)R.sup.41,
--NR.sup.40C(O)NR.sup.41R.sup.42,
--NR.sup.40S(O)R.sup.41--NR.sup.40S(O).sub.2R.sup.41,
--OP(O)R.sup.41R.sup.42, --P(O)R.sup.41R.sup.42,
--P(O)OR.sup.41R.sup.42, --P(O)(OR.sup.41)OR.sup.42,
--C(O)NR.sup.41R.sup.42, optionally substituted C.sub.1-6 alkyl,
optionally substituted C.sub.2-8 alkenyl, optionally substituted
C.sub.2-6 alkynyl, optionally substituted C.sub.3-6 cycloalkyl,
optionally substituted C.sub.6-12 arylalkyl, optionally substituted
C.sub.6-12 aryl, optionally substituted 3-14 membered heteroaryl,
optionally substituted C.sub.1-6 alkyloxy, optionally substituted
C.sub.2-6 alkenyloxy, optionally substituted C.sub.2-6 alkynyloxy,
optionally substituted C.sub.3-6 cycloalkyloxy, optionally
substituted C.sub.6-12 aryloxy, optionally substituted 3-14
membered heteroaryloxy, optionally substituted 4-12 membered
heterocyclyloxy, optionally substituted --C(O)C.sub.1-6 alkyl,
optionally substituted --C(O)C.sub.2-6 alkenyl, optionally
substituted --C(O)C.sub.2-6 alkynyl, optionally substituted
--C(O)C.sub.3-6 cycloalkyl, optionally substituted --C(O)C.sub.6-12
aryl, optionally substituted --C(O)-3-14 membered heteroaryl,
optionally substituted --C(O)C.sub.6-12 arylalkyl, optionally
substituted 3-10 membered heterocyclyl, --OH, --NR.sup.41R.sup.42,
--C(O)OR.sup.40, --CN, --N.sub.3,
--C(.dbd.NR.sup.43)NR.sup.41R.sup.42, --C(.dbd.NR.sup.43)OR.sup.40,
--NR.sup.40C(.dbd.NR.sup.43)NR.sup.41R.sup.42,
--NR.sup.41C(O)OR.sup.40, and --OC(O)NR.sup.41R.sup.42;
[0036] each R.sup.40, R.sup.41, and R.sup.42, independently is
selected from the group consisting of H, optionally substituted
C.sub.1-12 alkyl, optionally substituted C.sub.2-12 alkenyl,
optionally substituted C.sub.2-12 alkynyl, optionally substituted
C.sub.3-12 cycloalkyl, optionally substituted C.sub.6-14 aryl,
optionally substituted 3-14 membered heteroaryl, optionally
substituted 3-12 membered heterocyclyl, optionally substituted 3-18
membered heteroarylalkyl, and optionally substituted C.sub.6-18
arylalkyl;
[0037] or R.sup.41 and R.sup.42 taken together with the atoms to
which they are attached form a 3 to 10 membered heterocyclyl;
[0038] each R.sup.43, independently, is selected from the group
consisting of H, optionally substituted C.sub.1-12 alkyl,
optionally substituted C.sub.2-12 alkenyl, optionally substituted
C.sub.2-12 alkynyl, optionally substituted C.sub.3-12 cycloalkyl,
optionally substituted C.sub.6-14 aryl, optionally substituted 3-14
membered heteroaryl, optionally substituted 3-12 membered
heterocyclyl, optionally substituted 3-18 membered heteroarylalkyl,
optionally substituted C.sub.6-18 arylalkyl, --CN, --C(O)R.sup.44,
--CHO and --S(O).sub.2R.sup.44;
[0039] each R.sup.44 individually is optionally substituted
C.sub.1-12 alkyl;
[0040] wherein, each substituted Q.sup.4, substituted R.sup.40,
substituted R.sup.41, substituted R.sup.42, substituted R.sup.43,
or substituted R.sup.44 is independently substituted with one or
more Q.sup.5;
[0041] each Q.sup.5, individually, is selected from the group
consisting of halogen, oxo, oxide, --NO.sub.2, --N(.dbd.O),
--SR.sup.50, --S(O)R.sup.50, --S(O).sub.2R.sup.50,
--S(O).sub.2NR.sup.50R.sup.51, --NR.sup.50C(O)R.sup.51,
--NR.sup.50C(O)NR.sup.51R.sup.52, --NR.sup.50S(O)R.sup.51,
--NR.sup.50S(O).sub.2R.sup.51, --OP(O)R.sup.51R.sup.52,
--P(O)R.sup.51R.sup.52, --P(O)OR.sup.51R.sup.52,
--P(O)(OR.sup.51)OR.sup.52, --C(O)NR.sup.51R.sup.52, optionally
substituted C.sub.1-6 alkyl, optionally substituted C.sub.2-6
alkenyl, optionally substituted C.sub.2-6 alkynyl, optionally
substituted C.sub.3-6 cycloalkyl, optionally substituted C.sub.6-12
arylalkyl, optionally substituted C.sub.6-12 aryl, optionally
substituted 3-14 membered heteroaryl, optionally substituted
C.sub.1-6 alkyloxy, optionally substituted C.sub.2-6 alkenyloxy,
optionally substituted C.sub.2-6 alkynyloxy, optionally substituted
C.sub.3-6 cycloalkyloxy, optionally substituted C.sub.6-12 aryloxy,
optionally substituted 3-14 membered heteroaryloxy, optionally
substituted 4-12 membered heterocyclyloxy, optionally substituted
--C(O)C.sub.1-6 alkyl, optionally substituted --C(O)C.sub.2-6
alkenyl, optionally substituted --C(O)C.sub.2-6 alkynyl, optionally
substituted --C(O)C.sub.3-6 cycloalkyl, optionally substituted
--C(O)C.sub.6-12 aryl, optionally substituted --C(O)-3-14 membered
heteroaryl, optionally substituted --C(O)C.sub.6-12 arylalkyl,
optionally substituted 3-10 membered heterocyclyl, --OH,
--NR.sup.51R.sup.52, --C(O)OR.sup.50, --CN, --N.sub.3,
--C(.dbd.NR.sup.53)NR.sup.51R.sup.52, --C(.dbd.NR.sup.53)OR.sup.50,
--NR.sup.50C(.dbd.NR.sup.53)NR.sup.51R.sup.52,
--NR.sup.51C(O)OR.sup.50, and --OC(O)NR.sup.51R.sup.52;
[0042] each R.sup.50, R.sup.51, and R.sup.52, independently is
selected from the group consisting of H, optionally substituted
C.sub.1-12 alkyl, optionally substituted C.sub.2-12 alkenyl,
optionally substituted C.sub.2-12 alkynyl, optionally substituted
C.sub.3-12 cycloalkyl, optionally substituted C.sub.6-14 aryl,
optionally substituted 3-14 membered heteroaryl, optionally
substituted 3-12 membered heterocyclyl, optionally substituted 3-18
membered heteroarylalkyl, and optionally substituted C.sub.6-18
arylalkyl;
[0043] or R.sup.51 and R.sup.52 taken together with the atoms to
which they are attached form a 3 to 10 membered heterocyclyl;
[0044] each R.sup.53, independently, is selected from the group
consisting of H, optionally substituted C.sub.1-12 alkyl,
optionally substituted C.sub.2-12 alkenyl, optionally substituted
C.sub.2-12 alkynyl, optionally substituted C.sub.3-12 cycloalkyl,
optionally substituted C.sub.6-14 aryl, optionally substituted 3-14
membered heteroaryl, optionally substituted 3-12 membered
heterocyclyl, optionally substituted 3-18 membered heteroarylalkyl,
optionally substituted C.sub.6-18 arylalkyl, --CN, --C(O)R.sup.54,
--CHO and --S(O).sub.2R.sup.54;
[0045] each R.sup.54, independently, is optionally substituted
C.sub.1-12 alkyl;
[0046] wherein, each substituted Q.sup.5, substituted R.sup.50,
substituted R.sup.51, substituted R.sup.52, substituted R.sup.53,
or substituted R.sup.54 is independently substituted with one or
more Q.sup.6;
[0047] each Q.sup.6, independently, is selected from the group
consisting of halogen, oxo, oxide, --NO.sub.2, --N(.dbd.O),
--SR.sup.60, --S(O)R.sup.60, --S(O).sub.2R.sup.60,
--S(O).sub.2NR.sup.60R.sup.61, --NR.sup.60C(O)R.sup.61,
--NR.sup.60C(O)NR.sup.61R.sup.62, --NR.sup.60S(O)R.sup.61,
--NR.sup.60S(O).sub.2R.sup.61, --OP(O)R.sup.61R.sup.62,
--P(O)R.sup.61R.sup.62, --P(O)OR.sup.61R.sup.62,
--P(O)(OR.sup.61)OR.sup.62, --C(O)NR.sup.61R.sup.62, C.sub.1-6
alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-6 cycloalkyl,
C.sub.6-12 arylalkyl, C.sub.6-12 aryl, 3-14 membered heteroaryl,
C.sub.1-6 alkyloxy, C.sub.2-6 alkenyloxy, C.sub.2-6 alkynyloxy,
C.sub.3-6 cycloalkyloxy, C.sub.6-12 aryloxy, 3-14 membered
heteroaryloxy, 4-12 membered heterocyclyloxy, --C(O)C.sub.1-6
alkyl, --C(O)C.sub.2-6 alkenyl, --C(O)C.sub.2-6 alkynyl,
--C(O)C.sub.3-6 cycloalkyl, --C(O)C.sub.1-6 haloalkyl,
--C(O)C.sub.6-12 aryl, --C(O)-3-14 membered heteroaryl,
--C(O)C.sub.6-12 arylalkyl, 3-10 membered heterocyclyl, --OH,
--NR.sup.61R.sup.62, --C(O)OR.sup.60, --CN, --N.sub.3,
--C(.dbd.NR.sup.63)NR.sup.61R.sup.62, --C(.dbd.NR.sup.63)OR.sup.60,
--NR.sup.60C(.dbd.NR.sup.63)NR.sup.61R.sup.62,
--NR.sup.61C(O)OR.sup.60, and --OC(O)NR.sup.61R.sup.62;
[0048] each R.sup.60, R.sup.61, and R.sup.62, independently, is
selected from the group consisting of H, C.sub.1-12 alkyl,
C.sub.2-12 alkenyl, C.sub.2-12 alkynyl, C.sub.3-12 cycloalkyl,
C.sub.1-12 haloalkyl, C.sub.6-14 aryl, 3-14 membered heteroaryl,
3-12 membered heterocyclyl, 3-18 membered heteroarylalkyl, and
C.sub.6-18 arylalkyl;
[0049] or R.sup.61 and R.sup.62 taken together with the atoms to
which they are attached form a 3 to 10 membered heterocyclyl;
[0050] each R.sup.63 independently is selected from the group
consisting of H, C.sub.1-12 alkyl, C.sub.2-12 alkenyl, C.sub.2-12
alkynyl, C.sub.3-12 cycloalkyl, C.sub.6-14 aryl, 3-14 membered
heteroaryl, 3-12 membered heterocyclyl, 3-18 membered
heteroarylalkyl, C.sub.6-18 arylalkyl, --CN, --C(O)R.sup.64, --CHO
and --S(O).sub.2R.sup.64;
[0051] each R.sup.64 individually is C.sub.1-12 alkyl;
[0052] each R.sup.4 is independently H, C.sub.1-C.sub.12 alkyl,
C.sub.3-C.sub.12 cycloalkyl, 3-14 membered heteroaryl or 3-12
membered heterocyclyl wherein each C.sub.1-C.sub.12 alkyl,
C.sub.3-C.sub.12 cycloalkyl, 3-14 membered heteroaryl or 3-12
membered heterocyclyl is optionally substituted with one or more
Q.sup.1;
[0053] each R.sup.5 is independently C.sub.1-C.sub.12 alkyl,
C.sub.3-C.sub.12 cycloalkyl, 3-14 membered heteroaryl or 3-12
membered heterocyclyl wherein each C.sub.1-C.sub.12 alkyl,
C.sub.3-C.sub.12 cycloalkyl, 3-14 membered heteroaryl or 3-12
membered heterocyclyl is optionally substituted with one or more
Q.sup.1;
[0054] R.sup.6 is C.sub.1-C.sub.12 alkylyne, C.sub.3-C.sub.12
cycloalkylyne, or 3-12 membered heterocyclylyne wherein each
C.sub.1-C.sub.12 alkylyne, C.sub.3-C.sub.12 cycloalkylyne, or 3-12
membered heterocyclylyne is optionally substituted with one or more
Q.sup.1; and
[0055] R.sup.7 is selected from the group consisting of optionally
substituted C.sub.1-12 alkyl, optionally substituted C.sub.2-12
alkenyl, optionally substituted C.sub.2-12 alkynyl, optionally
substituted C.sub.3-12 cycloalkyl, optionally substituted
C.sub.6-14 aryl, optionally substituted 3-14 membered heteroaryl,
optionally substituted 3-12 membered heterocyclyl, optionally
substituted 3-18 membered heteroarylalkyl, optionally substituted
3-18 membered heterocyclylalkyl and optionally substituted
C.sub.6-18 arylalkyl;
[0056] wherein, each substituted R.sup.7 is substituted with one or
more Q.sup.2.
[0057] In another embodiment, a method for treating Flaviviridae
viral infections is provided comprising administering an effective
amount of a compound of Formula I to a patient in need thereof. The
compound of Formula I is administered to a human subject in need
thereof, such as a human being who is infected with viruses of the
Flaviviridae family. In another embodiment, the compound of Formula
I is administered to a human subject in need thereof, such as a
human being who is infected with a HCV virus. In one embodiment,
the treatment results in the reduction of one or more of the in
viral loads or clearance of RNA in the patient.
[0058] In another embodiment, provided is a method of treating
and/or preventing a disease caused by a viral infection wherein the
viral infection is caused by a virus selected from the group
consisting of dengue virus, yellow fever virus, West Nile virus,
Japanese encephalitis virus, tick-borne encephalitis virus, Junjin
virus, Murray Valley encephalitis virus, St Louis encephalitis
virus, Omsk hemorrhagic fever virus, bovine viral disarrhea virus,
Zika virus and Hepatitis C virus; by administering to a subject in
need thereof a therapeutically effective amount of a compound of
Formula I, or a pharmaceutically acceptable salt thereof.
[0059] In another embodiment, provided is the use of a compound of
Formula I for the manufacture of a medicament for the treatment of
Flaviviridae viral infections. In another aspect of this
embodiment, the Flaviviridae viral infection is an HCV
infection.
[0060] In another embodiment, provided is a compound of Formula I
for use in treating a Flaviviridae viral infection. In another
aspect of this embodiment, the Flaviviridae viral infection is an
acute or chronic HCV infection. In another aspect of this
embodiment, the treatment results in the reduction of one or more
of the viral loads or clearance of RNA in the patient. In another
aspect of this embodiment, the treatment results in the reduction
of the HCV viral load or clearance of HCV viral RNA in the
patient.
[0061] In another embodiment, provided is a method for treating or
preventing HCV comprising administering an effective amount of a
compound of Formula I to a patient in need thereof. In another
embodiment, provided is the use of a compound of the present
invention for the manufacture of a medicament for the treatment or
prevention of HCV.
[0062] In another embodiment, provided is a pharmaceutical
composition comprising a compound of Formula I and one or more
pharmaceutically acceptable carriers or excipients. The
pharmaceutical composition of Formula I may further comprise one or
more additional therapeutic agents. The one or more additional
therapeutic agent may be, without limitation, selected from:
interferons, ribavirin or its analogs, HCV NS3 protease inhibitors,
alpha-glucosidase 1 inhibitors, hepatoprotectants, nucleoside or
nucleotide inhibitors of HCV NS5B polymerase, non-nucleoside
inhibitors of HCV NS5B polymerase, HCV NS5A inhibitors, TLR-7
agonists, cyclophilin inhibitors, HCV IRES inhibitors,
pharmacokinetic enhancers, and other drugs for treating HCV, or
mixtures thereof.
[0063] In another embodiment, provided is a method for the
treatment or prevention of the symptoms or effects of an HCV
infection in an infected animal which comprises administering to,
i.e. treating, said animal with a pharmaceutical combination
composition or formulation comprising an effective amount of a
Formula I compound, and a second compound having anti-HCV
properties.
[0064] In another embodiment, provided are compounds of Formula I
and pharmaceutically acceptable salts thereof and all racemates,
enantiomers, diastereomers, tautomers, polymorphs, pseudopolymorphs
and amorphous forms thereof.
[0065] In another embodiment, provided are processes and novel
intermediates disclosed herein which are useful for preparing
Formula I compounds.
[0066] In other embodiments, novel methods for synthesis, analysis,
separation, isolation, purification, characterization, and testing
of the compounds of Formula I are provided.
[0067] The present invention includes combinations of aspects and
embodiments, as well as preferences, as herein described throughout
the present specification.
DETAILED DESCRIPTION
[0068] Reference will now be made in detail to certain embodiments
of the invention, examples of which are illustrated in the
accompanying structures and formulas. While the invention will be
described in conjunction with the enumerated embodiments, it will
be understood that they are not intended to limit the invention to
those embodiments. On the contrary, the invention is intended to
cover all alternatives, modifications, and equivalents, which may
be included within the scope of the present invention as defined by
the claims.
[0069] All documents referenced herein are each incorporated by
reference in their entirety for all purposes.
[0070] In one embodiment of Formula I, R.sup.1 is optionally
substituted C.sub.1-12 alkyl, optionally substituted C.sub.2-12
alkenyl, optionally substituted C.sub.2-12 alkynyl, or optionally
substituted C.sub.3-12 cycloalkyl. In another aspect of this
embodiment, R.sup.1 is optionally substituted C.sub.1-C.sub.12
alkyl. In another aspect of this embodiment, R.sup.1 is optionally
substituted C.sub.3-C.sub.7 secondary or tertiary alkyl. In another
aspect of this embodiment, R.sup.1 is prop-2-yl(isopropyl) or
2-methylprop-2-yl (t-butyl).
[0071] In another embodiment of Formula I, R.sup.2 is optionally
substituted C.sub.1-12 alkyl, optionally substituted C.sub.2-12
alkenyl, optionally substituted C.sub.2-12 alkynyl, optionally
substituted C.sub.3-12 cycloalkyl, or optionally substituted
C.sub.6-18 arylalkyl. In another aspect of this embodiment, R.sup.2
is optionally substituted C.sub.3-12 cycloalkyl. In another aspect
of this embodiment, R.sup.2 is optionally substituted
methylcyclohexyl or optionally substituted methylcyclohexenyl. In
another aspect of this embodiment, R.sup.2 is optionally
substituted 4-methylcyclohexyl. In another aspect of this
embodiment, R.sup.2 is optionally substituted 4-methylcyclohexenyl.
In a preferred aspect of this embodiment, R.sup.2 is
##STR00003##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00004##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00005##
[0072] In another preferred aspect of this embodiment, R.sup.2
is
##STR00006##
[0073] In another embodiment of Formula I, Y is --R.sup.3-L-Het. In
another embodiment of Formula I, Y is --N(R.sup.4)(R.sup.5). In
another embodiment of Formula I, Y is --R.sup.6.dbd.NOR.sup.7.
[0074] In another embodiment of Formula I, Y is --R.sup.3-L-Het,
R.sup.1 is optionally substituted C.sub.1-C.sub.12 alkyl, and
R.sup.2 is optionally substituted C.sub.3-12 cycloalkyl. In another
aspect of this embodiment, R.sup.3 is optionally substituted
C.sub.1-12 alkylene, C.sub.2-12 alkenylene, substituted C.sub.2-12
alkenylene, C.sub.2-12 alkynylene, substituted C.sub.2-12
alkynylene, C.sub.3-12 cycloalkylene, substituted C.sub.3-12
cycloalkylene, C.sub.3-12 cycloalkylalkylene, substituted
C.sub.3-12 cycloalkylalkylene, optionally substituted C.sub.6-14
arylene, optionally substituted 3-14 membered heteroarylene,
optionally substituted 3-12 membered heterocyclylene, optionally
substituted 3-18 membered heteroarylalkylene, or optionally
substituted C.sub.6-18 arylalkylene. In another aspect of this
embodiment, R.sup.3 is optionally substituted C.sub.1-12 alkylene,
C.sub.3-12 cycloalkylene, substituted C.sub.3-12 cycloalkylene,
C.sub.3-12 cycloalkylalkylene, substituted C.sub.3-12
cycloalkylalkylene, optionally substituted C.sub.6-14 arylene, or
optionally substituted 3-12 membered heterocyclylene. In another
aspect of this embodiment, L is --OC(O)N(R.sup.4)--,
--N(R.sup.4)C(O)O--, --N(R.sup.4)S(O).sub.2--, --N(R.sup.4)C(O)--,
--C(O)--, --C(O)O--, --OC(O)--, --N(R.sup.4)N(R.sup.4)C(O)O--, or
--N(R.sup.4)N(R.sup.4)--. In another aspect of this embodiment, L
is --OC(O)N(R.sup.4)--. In another aspect of this embodiment, L is
--N(R.sup.4)C(O)O--. In another aspect of this embodiment, L is
--N(R.sup.4)S(O).sub.2--. In another aspect of this embodiment, L
is --N(R.sup.4)C(O)--. In another aspect of this embodiment, L is
--C(O)--. In another aspect of this embodiment, L is --C(O)O--. In
another aspect of this embodiment, L is --OC(O)--. In another
aspect of this embodiment, L is --N(R.sup.4)N(R.sup.4)C(O)O--. In
another aspect of this embodiment, L is
--N(R.sup.4)N(R.sup.4)--.
[0075] In another embodiment of Formula I, Y is --R.sup.3-L-Het,
R.sup.1 is optionally substituted C.sub.1-C.sub.12 alkyl, R.sup.2
is optionally substituted C.sub.3-12 cycloalkyl and R.sup.3 is
optionally substituted C.sub.1-12 alkylene, C.sub.3-12
cycloalkylene, substituted C.sub.3-12 cycloalkylene, C.sub.3-12
cycloalkylalkylene, substituted C.sub.3-12 cycloalkylalkylene,
optionally substituted C.sub.6-14 arylene, or optionally
substituted 3-12 membered heterocyclylene. In another aspect of
this embodiment, R.sup.3 is optionally substituted C.sub.1-6
alkylene. In another aspect of this embodiment, R.sup.3 is
C.sub.4-6 cycloalkylene or substituted C.sub.4-6 cycloalkylene. In
another aspect of this embodiment, R.sup.3 is an optionally
substituted 5-6 membered heterocyclylene. In another aspect of this
embodiment, L is --OC(O)N(R.sup.4)--, --N(R.sup.4)C(O)O--,
--N(R.sup.4)S(O).sub.2--, --N(R.sup.4)C(O)--, --C(O)--, --C(O)O--,
--OC(O)--, --N(R.sup.4)N(R.sup.4)C(O)O--, or
--N(R.sup.4)N(R.sup.4)--. In another aspect of this embodiment, L
is --OC(O)N(R.sup.4)--. In another aspect of this embodiment, L is
--N(R.sup.4)C(O)O--. In another aspect of this embodiment,
--N(R.sup.4)S(O).sub.2--. In another aspect of this embodiment, L
is --N(R.sup.4)C(O)--. In another aspect of this embodiment, L is
--C(O)--. In another aspect of this embodiment, L is --C(O)O--. In
another aspect of this embodiment, L is --OC(O)--. In another
aspect of this embodiment, L is --N(R.sup.4)N(R.sup.4)C(O)O--. In
another aspect of this embodiment, L is --N(R.sup.4)N(R.sup.4)--.
In another aspect of this embodiment, Het is an optionally
substituted 3-12 membered heterocyclyl or optionally substituted
3-14 membered heteroaryl wherein said optionally substituted 3-12
membered heterocyclyl or optionally substituted 3-14 membered
heteroaryl comprises one to four heteroatoms selected from O, S, or
N. In another aspect of this embodiment, Het is an optionally
substituted 3-12 membered heterocyclyl comprising one or two
heteroatoms selected from O, S, or N. In another aspect of this
embodiment, Het is an optionally substituted 5-10 membered
heteroaryl comprising one to four heteroatoms selected from O, S,
or N.
[0076] In another embodiment of Formula I, Y is
--N(R.sup.4)(R.sup.5), R.sup.1 is optionally substituted
C.sub.1-C.sub.12 alkyl, and R.sup.2 is optionally substituted
C.sub.3-12 cycloalkyl. In another aspect of this embodiment,
R.sup.4 is independently H or C.sub.1-C.sub.12 alkyl wherein
C.sub.1-C.sub.12 alkyl is optionally substituted with one or more
Q.sup.1. In another aspect of this embodiment, R.sup.4 is H or
optionally substituted C.sub.1-C.sub.6 alkyl. In another aspect of
this embodiment, R.sup.5 is C.sub.1-C.sub.12 alkyl,
C.sub.3-C.sub.12 cycloalkyl, 3-14 membered heteroaryl or 3-12
membered heterocyclyl wherein each C.sub.1-C.sub.12 alkyl,
C.sub.3-C.sub.12 cycloalkyl, 3-14 membered heteroaryl or 3-12
membered heterocyclyl is optionally substituted with one or more
Q.sup.1. In another aspect of this embodiment, R.sup.5 is
C.sub.1-C.sub.6 alkyl, C.sub.5-C.sub.6 cycloalkyl, 5-10 membered
heteroaryl or 4-10 membered heterocyclyl wherein each
C.sub.1-C.sub.6 alkyl, C.sub.5-C.sub.6 cycloalkyl, 5-10 membered
heteroaryl or 4-10 membered heterocyclyl is optionally substituted
with one or more Q.sup.1.
[0077] In another embodiment of Formula I, Y is
--R.sup.6.dbd.NOR.sup.7, R.sup.1 is optionally substituted
C.sub.1-C.sub.12 alkyl, and R.sup.2 is optionally substituted
C.sub.3-12 cycloalkyl. In another aspect of this embodiment,
R.sup.6 is C.sub.1-C.sub.12 alkylyne, C.sub.3-C.sub.12
cycloalkylyne, or 3-12 membered heterocyclylyne wherein each
C.sub.1-C.sub.12 alkylyne, C.sub.3-C.sub.12 cycloalkylyne, or 3-12
membered heterocyclylyne is optionally substituted with one or more
Q. In another aspect of this embodiment, R.sup.6 is C.sub.5-C.sub.6
cycloalkylyne or 4-6 membered heterocyclylyne wherein each
C.sub.5-C.sub.6 cycloalkylyne or 4-6 membered heterocyclylyne is
optionally substituted with one or more Q.sup.1. In another aspect
of this embodiment, R.sup.6 is cyclohexylyne. In another aspect of
this embodiment, R.sup.7 is optionally substituted C.sub.1-12
alkyl, optionally substituted C.sub.2-12 alkenyl, optionally
substituted C.sub.2-12 alkynyl, optionally substituted C.sub.3-12
cycloalkyl, optionally substituted C.sub.6-14 aryl, optionally
substituted 3-14 membered heteroaryl, optionally substituted 3-12
membered heterocyclyl, optionally substituted 3-18 membered
heteroarylalkyl, optionally substituted 3-18 membered
heterocyclylalkyl or optionally substituted C.sub.6-18 arylalkyl;
wherein, when R.sup.7 is substituted, R.sup.7 is substituted with
one or more Q.sup.2. In another aspect of this embodiment, R.sup.7
is optionally substituted C.sub.1-C.sub.6 alkyl. In another aspect
of this embodiment, R.sup.7 is optionally substituted
C.sub.7-C.sub.11 arylalkyl. In another aspect of this embodiment,
R.sup.7 is optionally substituted 6-11 membered heteroarylalkyl. In
another aspect of this embodiment, R.sup.7 is optionally
substituted 6-11 membered heterocyclylalkyl.
[0078] In another embodiment of Formula I, R.sup.1 is optionally
substituted C.sub.3-C.sub.7 secondary or tertiary alkyl and R.sup.2
is optionally substituted methylcyclohexyl or optionally
substituted methylcyclohexenyl. In another aspect of this
embodiment, R.sup.1 is prop-2-yl (isopropyl) or 2-methylprop-2-yl
(t-butyl). In another aspect of this embodiment, R.sup.2 is
optionally substituted 4-methylcyclohexyl. In a preferred aspect of
this embodiment, R.sup.2 is
##STR00007##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00008##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00009##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00010##
[0079] In another embodiment of Formula I, Y is --R.sup.3-L-Het,
R.sup.1 is optionally substituted C.sub.3-C.sub.7 secondary or
tertiary alkyl and R.sup.2 is optionally substituted
methylcyclohexyl or optionally substituted methylcyclohexenyl. In
another aspect of this embodiment, R.sup.3 is optionally
substituted C.sub.1-6 alkylene. In another aspect of this
embodiment, R.sup.3 is C.sub.4-6 cycloalkylene or substituted
C.sub.4-6 cycloalkylene. In another aspect of this embodiment,
R.sup.3 is an optionally substituted 5-6 membered heterocyclylene.
In another aspect of this embodiment, L is --OC(O)N(R.sup.4)--,
--N(R.sup.4)C(O)O--, --N(R.sup.4)S(O).sub.2--, --N(R.sup.4)C(O)--,
--C(O)--, --C(O)O--, --OC(O)--, --N(R.sup.4)N(R.sup.4)C(O)O--, or
--N(R.sup.4)N(R.sup.4)--. In another aspect of this embodiment, L
is --OC(O)N(R.sup.4)--. In another aspect of this embodiment, L is
--N(R.sup.4)C(O)O--. In another aspect of this embodiment, L is
--N(R.sup.4)S(O).sub.2--. In another aspect of this embodiment, L
is --N(R.sup.4)C(O)--. In another aspect of this embodiment, L is
--C(O)--. In another aspect of this embodiment, --C(O)O--. In
another aspect of this embodiment, L is --OC(O)--. In another
aspect of this embodiment, L is --N(R.sup.4)N(R.sup.4)C(O)O--. In
another aspect of this embodiment, L is --N(R.sup.4)N(R.sup.4)--.
In another aspect of this embodiment, Het is an optionally
substituted 3-12 membered heterocyclyl or optionally substituted
3-14 membered heteroaryl wherein said optionally substituted 3-12
membered heterocyclyl or optionally substituted 3-14 membered
heteroaryl comprises one to four heteroatoms selected from O, S, or
N. In another aspect of this embodiment, Het is an optionally
substituted 3-12 membered heterocyclyl comprising one or two
heteroatoms selected from O, S, or N. In aspect of this embodiment,
Het is an optionally substituted 5-10 membered heteroaryl
comprising one to four heteroatoms selected from O, S, or N.
[0080] In another embodiment of Formula I, Y is
--R.sup.3--N(R.sup.4)C(O)O-Het, R.sup.1 is optionally substituted
C.sub.3-C.sub.7 secondary or tertiary alkyl and R.sup.2 is
optionally substituted methylcyclohexyl or optionally substituted
methylcyclohexenyl. In another aspect of this embodiment, R.sup.3
is optionally substituted C.sub.1-6 alkylene. In another aspect of
this embodiment, R.sup.3 is C.sub.4-6 cycloalkylene or substituted
C.sub.4-6 cycloalkylene. In another aspect of this embodiment, Het
is an optionally substituted 3-12 membered heterocyclyl or
optionally substituted 3-14 membered heteroaryl wherein said
optionally substituted 3-12 membered heterocyclyl or optionally
substituted 3-14 membered heteroaryl comprises one to four
heteroatoms selected from O, S, or N. In another aspect of this
embodiment, Het is an optionally substituted 3-12 membered
heterocyclyl comprising one or two heteroatoms selected from O, S,
or N. In aspect of this embodiment, Het is an optionally
substituted 5-10 membered heteroaryl comprising one to four
heteroatoms selected from O, S, or N.
[0081] In another embodiment of Formula I, Y is
--R.sup.3--C(O)O-Het, R.sup.1 is optionally substituted
C.sub.3-C.sub.7 secondary or tertiary alkyl, R.sup.2 is optionally
substituted methylcyclohexyl or optionally substituted
methylcyclohexenyl and R.sup.3 is an optionally substituted 5-6
membered heterocyclylene. In another aspect of this embodiment, Het
is an optionally substituted 3-12 membered heterocyclyl or
optionally substituted 3-14 membered heteroaryl wherein said
optionally substituted 3-12 membered heterocyclyl or optionally
substituted 3-14 membered heteroaryl comprises one to four
heteroatoms selected from O, S, or N. In another aspect of this
embodiment, Het is an optionally substituted 3-12 membered
heterocyclyl comprising one or two heteroatoms selected from O, S,
or N. In aspect of this embodiment, Het is an optionally
substituted 5-10 membered heteroaryl comprising one to four
heteroatoms selected from O, S, or N. In another aspect of this
embodiment --R.sup.3--C(O)O-Het is:
##STR00011##
wherein the pyrrolidinyl or piperidinyl ring is optionally
substituted.
[0082] In another embodiment of Formula I, Y is
--N(R.sup.4)(R.sup.5), R.sup.1 is optionally substituted
C.sub.3-C.sub.7 secondary or tertiary alkyl and R.sup.2 is
optionally substituted methylcyclohexyl or optionally substituted
methylcyclohexenyl. In another aspect of this embodiment, R.sup.4
is H or optionally substituted C.sub.1-C.sub.6 alkyl. In another
aspect of this embodiment, R.sup.5 is C.sub.1-C.sub.12 alkyl,
C.sub.3-C.sub.12 cycloalkyl, 3-14 membered heteroaryl or 3-12
membered heterocyclyl wherein each C.sub.1-C.sub.12 alkyl,
C.sub.3-C.sub.12 cycloalkyl, 3-14 membered heteroaryl or 3-12
membered heterocyclyl is optionally substituted with one or more
Q.sup.1. In another aspect of this embodiment, R.sup.5 is
C.sub.1-C.sub.6 alkyl, C.sub.5-C.sub.6 cycloalkyl, 5-10 membered
heteroaryl or 4-10 membered heterocyclyl wherein each
C.sub.1-C.sub.6 alkyl, C.sub.5-C.sub.6 cycloalkyl, 5-10 membered
heteroaryl or 4-10 membered heterocyclyl is optionally substituted
with one or more Q.sup.1.
[0083] In another embodiment of Formula I, Y is
--R.sup.6.dbd.NOR.sup.7, R.sup.1 is optionally substituted
C.sub.3-C.sub.7 secondary or tertiary alkyl and R.sup.2 is
optionally substituted methylcyclohexyl or optionally substituted
methylcyclohexenyl. In another aspect of this embodiment, R.sup.6
is C.sub.5-C.sub.6 cycloalkylyne, or 4-6 membered heterocyclylyne
wherein each C.sub.5-C.sub.6 cycloalkylyne or 4-6 membered
heterocyclylyne is optionally substituted with one or more Q.sup.1.
In another aspect of this embodiment, R.sup.6 is cyclohexylyne. In
another aspect of this embodiment, R.sup.7 is optionally
substituted C.sub.1-C.sub.6 alkyl. In another aspect of this
embodiment, R.sup.7 is optionally substituted C.sub.7-C.sub.11
arylalkyl. In another aspect of this embodiment, R.sup.7 is
optionally substituted 6-11 membered heteroarylalkyl. In another
aspect of this embodiment, R.sup.7 is optionally substituted 6-11
membered heterocyclylalkyl.
[0084] In another embodiment, compounds of Formula I comprise
compounds of Formula II:
##STR00012##
or pharmaceutically acceptable salts and esters thereof, wherein:
R.sup.2 is optionally substituted 4-methylcyclohexyl or optionally
substituted methylcyclohexenyl and the remaining variables are
defined as for Formula I.
[0085] In one embodiment of Formula II, R.sup.2 is:
##STR00013##
[0086] In one embodiment of Formula II, R.sup.2 is:
##STR00014##
[0087] In another embodiment, R.sup.2 is
##STR00015##
[0088] In another embodiment, R.sup.2 is
##STR00016##
[0089] In one embodiment of Formula II, Y is --R.sup.3-L-Het. In
another aspect of this embodiment, R.sup.3 is optionally
substituted C.sub.1-5 alkylene. In another aspect of this
embodiment, R.sup.3 is C.sub.4-6 cycloalkylene or substituted
C.sub.4-6 cycloalkylene. In another aspect of this embodiment,
R.sup.3 is an optionally substituted 5-6 membered heterocyclylene.
In another aspect of this embodiment, L is --OC(O)N(R.sup.4)--,
N(R.sup.4)C(O)O--, --N(R.sup.4)S(O).sub.2--, --N(R.sup.4)C(O)--,
--C(O)--, --C(O)O--, --OC(O)--, --N(R.sup.4)N(R.sup.4)C(O)O--, or
--N(R.sup.4)N(R.sup.4)--. In another aspect of this embodiment, L
is --OC(O)N(R.sup.4)--. In another aspect of this embodiment, L is
--N(R.sup.4)C(O)O--. In another aspect of this embodiment, L is
--N(R.sup.4)S(O).sub.2--. In another aspect of this embodiment, L
is --N(R.sup.4)C(O)--. In another aspect of this embodiment,
--OC(O)--. In another aspect of this embodiment, --C(O)O--. In
another aspect of this embodiment, L is --OC(O)--. In another
aspect of this embodiment, L is --N(R.sup.4)N(R.sup.4)C(O)O--. In
another aspect of this embodiment, L is --N(R.sup.4)N(R.sup.4)--.
In another aspect of this embodiment, Het is an optionally
substituted 3-12 membered heterocyclyl or optionally substituted
3-14 membered heteroaryl wherein said optionally substituted 3-12
membered heterocyclyl or optionally substituted 3-14 membered
heteroaryl comprises one to four heteroatoms selected from O, S, or
N. In another aspect of this embodiment, Het is an optionally
substituted 3-12 membered heterocyclyl comprising one or two
heteroatoms selected from O, S, or N. In another aspect of this
embodiment, Het is an optionally substituted 5-10 membered
heteroaryl comprising one to four heteroatoms selected from O, S,
or N. In another aspect of this embodiment, R.sup.2 is:
##STR00017##
In another aspect of this embodiment, R.sup.2 is:
##STR00018##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00019##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00020##
[0090] In another embodiment of Formula II, Y is --R.sup.3-L-Het
wherein Het is an optionally substituted 3-12 membered heterocyclyl
or optionally substituted 3-14 membered heteroaryl wherein said
optionally substituted 3-12 membered heterocyclyl or optionally
substituted 3-14 membered heteroaryl comprises one to four
heteroatoms selected from O, S, or N. In another aspect of this
embodiment, Het is optionally substituted pyridinyl. In another
aspect of this embodiment, Het is optionally substituted
pyridazinyl. In another aspect of this embodiment, Het is
optionally substituted tetrahydro-2H-pyranyl. In another aspect of
this embodiment, Het is optionally substituted piperidinyl. In
another aspect of this embodiment, Het is optionally substituted
pyrrolidinyl. In another aspect of this embodiment, Het is
optionally substituted tetrahydrothiophenyl. In another aspect of
this embodiment, Het is optionally substituted pyrazinyl. In
another aspect of this embodiment, Het is optionally substituted
1H-tetrazolyl. In another aspect of this embodiment, Het is
optionally substituted azetidinyl. In another aspect of this
embodiment, Het is optionally substituted tetrahydrofuranyl. In
another aspect of this embodiment, Het is optionally substituted
tetrahydro-2H-furo[2,3-b]furanyl. In another aspect of this
embodiment, Het is optionally substituted thiazoyl. In another
aspect of this embodiment, Het is optionally substituted
1H-imidazolyl. In another aspect of this embodiment, Het is
optionally substituted 4H-1,2,4-triazolyl. In another aspect of
this embodiment, Het is optionally substituted 1H-pyrazolyl. In
another aspect of this embodiment, Het is optionally substituted
1,3,4-thiadiazolyl. In another aspect of this embodiment, Het is
optionally substituted quinolinyl. In another aspect of this
embodiment, Het is optionally substituted
[1,2,4]triazolo[4,3-a]pyridinyl. In another aspect of this
embodiment, Het is optionally substituted thiophenyl. In another
aspect of this embodiment, Het is optionally substituted
1,2,4-thiadiazolyl. In another aspect of this embodiment, Het is
optionally substituted pyrimidinyl. In another aspect of this
embodiment, Het is optionally substituted 1H-1,2,3-triazolyl. In
another aspect of this embodiment, Het is optionally substituted
1,3,4-oxadiazolyl. In another aspect of this embodiment, Het is
optionally substituted imidazo[1,2-b]pyridazinyl. In another aspect
of this embodiment, R.sup.2 is:
##STR00021##
In another aspect of this embodiment, R.sup.2 is:
##STR00022##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00023##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00024##
[0091] In another embodiment of Formula II, Y is
--R.sup.3--N(R.sup.4)C(O)O-Het. In another aspect of this
embodiment, R.sup.3 is optionally substituted C.sub.1-6 alkylene.
In another aspect of this embodiment, R.sup.3 is C.sub.4-6
cycloalkylene or substituted C.sub.4-6 cycloalkylene. In another
aspect of this embodiment, Het is an optionally substituted 3-12
membered heterocyclyl or optionally substituted 3-14 membered
heteroaryl wherein said optionally substituted 3-12 membered
heterocyclyl or optionally substituted 3-14 membered heteroaryl
comprises one to four heteroatoms selected from O, S, or N. In
another aspect of this embodiment, Het is an optionally substituted
3-12 membered heterocyclyl comprising one or two heteroatoms
selected from O, S, or N. In aspect of this embodiment, Het is an
optionally substituted 5-10 membered heteroaryl comprising one to
four heteroatoms selected from O, S, or N. In another aspect of
this embodiment, Het is optionally substituted pyridinyl. In
another aspect of this embodiment, Het is optionally substituted
pyridazinyl. In another aspect of this embodiment, Het is
optionally substituted tetrahydro-2H-pyranyl. In another aspect of
this embodiment, Het is optionally substituted piperidinyl. In
another aspect of this embodiment, Het is optionally substituted
pyrrolidinyl. In another aspect of this embodiment, Het is
optionally substituted tetrahydrothiophenyl. In another aspect of
this embodiment, Het is optionally substituted pyrazinyl. In
another aspect of this embodiment, Het is optionally substituted
1H-tetrazolyl. In another aspect of this embodiment, Het is
optionally substituted azetidinyl. In another aspect of this
embodiment, Het is optionally substituted tetrahydrofuranyl. In
another aspect of this embodiment, Het is optionally substituted
tetrahydro-2H-furo[2,3-b]furanyl. In another aspect of this
embodiment, Het is optionally substituted thiazoyl. In another
aspect of this embodiment, Het is optionally substituted
1H-imidazolyl. In another aspect of this embodiment, Het is
optionally substituted 4H-1,2,4-triazolyl. In another aspect of
this embodiment, Het is optionally substituted 1H-pyrazolyl. In
another aspect of this embodiment, Het is optionally substituted
1,3,4-thiadiazolyl. In another aspect of this embodiment, Het is
optionally substituted quinolinyl. In another aspect of this
embodiment, Het is optionally substituted
[1,2,4]triazolo[4,3-a]pyridinyl. In another aspect of this
embodiment, Het is optionally substituted thiophenyl. In another
aspect of this embodiment, Het is optionally substituted
1,2,4-thiadiazolyl. In another aspect of this embodiment, Het is
optionally substituted pyrimidinyl. In another aspect of this
embodiment, Het is optionally substituted 1H-1,2,3-triazolyl. In
another aspect of this embodiment, Het is optionally substituted
1,3,4-oxadiazolyl. In another aspect of this embodiment, Het is
optionally substituted imidazo[1,2-b]pyridazinyl. In another aspect
of this embodiment, R.sup.2 is:
##STR00025##
In another aspect of this embodiment, R.sup.2 is:
##STR00026##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00027##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00028##
[0092] In another embodiment of Formula II, Y is
--R.sup.3--C(O)O-Het and R.sup.3 is an optionally substituted 5-6
membered heterocyclylene. In another aspect of this embodiment, Het
is an optionally substituted 3-12 membered heterocyclyl or
optionally substituted 3-14 membered heteroaryl wherein said
optionally substituted 3-12 membered heterocyclyl or optionally
substituted 3-14 membered heteroaryl comprises one to four
heteroatoms selected from O, S, or N. In another aspect of this
embodiment, Het is an optionally substituted 3-12 membered
heterocyclyl comprising one or two heteroatoms selected from O, S,
or N. In aspect of this embodiment, Het is an optionally
substituted 5-10 membered heteroaryl comprising one to four
heteroatoms selected from O, S, or N. In another aspect of this
embodiment, Het is optionally substituted pyridinyl. In another
aspect of this embodiment, Het is optionally substituted
pyridazinyl. In another aspect of this embodiment, Het is
optionally substituted tetrahydro-2H-pyranyl. In another aspect of
this embodiment, Het is optionally substituted piperidinyl. In
another aspect of this embodiment, Het is optionally substituted
pyrrolidinyl. In another aspect of this embodiment, Het is
optionally substituted tetrahydrothiophenyl. In another aspect of
this embodiment, Het is optionally substituted pyrazinyl. In
another aspect of this embodiment, Het is optionally substituted
1H-tetrazolyl. In another aspect of this embodiment, Het is
optionally substituted azetidinyl. In another aspect of this
embodiment, Het is optionally substituted tetrahydrofuranyl. In
another aspect of this embodiment, Het is optionally substituted
tetrahydro-2H-furo[2,3-b]furanyl. In another aspect of this
embodiment, Het is optionally substituted thiazoyl. In another
aspect of this embodiment, Het is optionally substituted
1H-imidazolyl. In another aspect of this embodiment, Het is
optionally substituted 4H-1,2,4-triazolyl. In another aspect of
this embodiment, Het is optionally substituted 1H-pyrazolyl. In
another aspect of this embodiment, Het is optionally substituted
1,3,4-thiadiazolyl. In another aspect of this embodiment, Het is
optionally substituted quinolinyl. In another aspect of this
embodiment, Het is optionally substituted
[1,2,4]triazolo[4,3-a]pyridinyl. In another aspect of this
embodiment, Het is optionally substituted thiophenyl. In another
aspect of this embodiment, Het is optionally substituted
1,2,4-thiadiazolyl. In another aspect of this embodiment, Het is
optionally substituted pyrimidinyl. In another aspect of this
embodiment, Het is optionally substituted 1H-1,2,3-triazolyl. In
another aspect of this embodiment, Het is optionally substituted
1,3,4-oxadiazolyl. In another aspect of this embodiment, Het is
optionally substituted imidazo[1,2-b]pyridazinyl. In another aspect
of this embodiment --R.sup.3--C(O)O-Het is:
##STR00029##
wherein the pyrrolidinyl or piperidinyl ring is optionally
substituted. In another aspect of this embodiment, R.sup.2 is:
##STR00030##
In another aspect of this embodiment, R.sup.2 is:
##STR00031##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00032##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00033##
[0093] In another embodiment of Formula II, Y is
--N(R.sup.4)(R.sup.5). In another aspect of this embodiment,
R.sup.4 is H or optionally substituted C.sub.1-C.sub.6 alkyl. In
another aspect of this embodiment, R.sup.5 is C.sub.1-C.sub.6
alkyl, C.sub.5-C.sub.6 cycloalkyl, 5-10 membered heteroaryl or 4-10
membered heterocyclyl wherein each C.sub.1-C.sub.6 alkyl,
C.sub.5-C.sub.6 cycloalkyl, 5-10 membered heteroaryl or 4-10
membered heterocyclyl is optionally substituted with one or more
Q.sup.1. In another aspect of this embodiment, R.sup.4 is
optionally substituted C.sub.1-C.sub.6 alkyl and R.sup.5 is
C.sub.1-C.sub.6 alkyl, C.sub.5-C.sub.6 cycloalkyl, 5-10 membered
heteroaryl or 4-10 membered heterocyclyl wherein each
C.sub.1-C.sub.6 alkyl, C.sub.5-C.sub.6 cycloalkyl, 5-10 membered
heteroaryl or 4-10 membered heterocyclyl is optionally substituted
with one or more Q.sup.1. In another aspect of this embodiment,
R.sup.2 is:
##STR00034##
In another aspect of this embodiment, R.sup.2 is:
##STR00035##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00036##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00037##
[0094] In another embodiment of Formula II, Y is
--R.sup.6.dbd.NOR.sup.7. In another aspect of this embodiment,
R.sup.6 is C.sub.5-C.sub.6 cycloalkylyne, or 4-6 membered
heterocyclylyne wherein each C.sub.5-C.sub.6 cycloalkylyne or 4-6
membered heterocyclylyne is optionally substituted with one or more
Q. In another aspect of this embodiment, R.sup.6 is cyclohexylyne.
In another aspect of this embodiment, R.sup.7 is optionally
substituted C.sub.6 alkyl. In another aspect of this embodiment,
R.sup.7 is optionally substituted C.sub.7-C.sub.11 arylalkyl. In
another aspect of this embodiment, R.sup.7 is optionally
substituted 6-11 membered heteroarylalkyl. In another aspect of
this embodiment, R.sup.7 is optionally substituted 6-11 membered
heterocyclylalkyl. In another aspect of this embodiment, R.sup.6 is
cyclohexylyne and R.sup.7 is optionally substituted C.sub.1-C.sub.6
alkyl. In another aspect of this embodiment, R.sup.6 is
cyclohexylyne and R.sup.7 is optionally substituted
C.sub.7-C.sub.11 arylalkyl. In another aspect of this embodiment,
R.sup.6 is cyclohexylyne and R.sup.7 is optionally substituted 6-11
membered heteroarylalkyl. In another aspect of this embodiment,
R.sup.6 is cyclohexylyne and R.sup.7 is optionally substituted 6-11
membered heterocyclylalkyl.
In another aspect of this embodiment, R.sup.2 is:
##STR00038##
In another aspect of this embodiment, R.sup.2 is:
##STR00039##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00040##
In another preferred aspect of this embodiment, R.sup.2 is
##STR00041##
[0095] In another embodiment, the compound of Formula I or Formula
II is
##STR00042## ##STR00043## ##STR00044## ##STR00045## ##STR00046##
##STR00047## ##STR00048## ##STR00049## ##STR00050## ##STR00051##
##STR00052## ##STR00053## ##STR00054## ##STR00055##
##STR00056##
or a pharmaceutically acceptable salt or ester thereof.
DEFINITIONS
[0096] Unless stated otherwise, the following terms and phrases as
used herein are intended to have the following meanings. The fact
that a particular term or phrase is not specifically defined should
not be correlated to indefiniteness or lacking clarity, but rather
terms herein are used within their ordinary meaning. When trade
names are used herein, applicants intend to independently include
the tradename product and the active pharmaceutical ingredient(s)
of the tradename product.
[0097] The term "treating", and grammatical equivalents thereof,
when used in the context of treating a disease, means slowing or
stopping the progression of a disease, or ameliorating at least one
symptom of a disease, more preferably ameliorating more than one
symptom of a disease. For example, treatment of a hepatitis C virus
infection can include reducing the HCV viral load in an HCV
infected human being, and/or reducing the severity of jaundice
present in an HCV infected human being.
[0098] "Alkyl" is hydrocarbon containing normal, secondary,
tertiary or cyclic carbon atoms. For example, an alkyl group can
have 1 to 20 carbon atoms (i.e, C.sub.1-C.sub.20 alkyl), 1 to 10
carbon atoms (i.e., C.sub.1-C.sub.10 alkyl), or 1 to 6 carbon atoms
(i.e., C.sub.1-C.sub.6 alkyl). Examples of suitable alkyl groups
include, but are not limited to, methyl (Me, --CH.sub.3), ethyl
(Et, --CH.sub.2CH.sub.3), 1-propyl (n-Pr, n-propyl,
--CH.sub.2CH.sub.2CH.sub.3), 2-propyl (i-Pr, i-propyl,
--CH(CH.sub.3).sub.2), 1-butyl (n-Bu, n-butyl,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-methyl-1-propyl (i-Bu,
i-butyl, --CH.sub.2CH(CH.sub.3).sub.2), 2-butyl (s-Bu, s-butyl,
--CH(CH.sub.3)CH.sub.2CH.sub.3), 2-methyl-2-propyl (t-Bu, t-butyl,
--C(CH.sub.3).sub.3), 1-pentyl (n-pentyl,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-pentyl
(--CH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.3), 3-pentyl
(--CH(CH.sub.2CH.sub.3).sub.2), 2-methyl-2-butyl
(--C(CH.sub.3).sub.2CH.sub.2CH.sub.3), 3-methyl-2-butyl
(--CH(CH.sub.3)CH(CH.sub.3).sub.2), 3-methyl-1-butyl
(--CH.sub.2CH.sub.2CH(CH.sub.3).sub.2), 2-methyl-1-butyl
(--CH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.3), 1-hexyl
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-hexyl
(--CH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 3-hexyl
(--CH(CH.sub.2CH.sub.3)(CH.sub.2CH.sub.2CH.sub.3)),
2-methyl-2-pentyl (--C(CH.sub.3).sub.2CH.sub.2CH.sub.2CH.sub.3),
3-methyl-2-pentyl (--CH(CH.sub.3)CH(CH.sub.3)CH.sub.2CH.sub.3),
4-methyl-2-pentyl (--CH(CH.sub.3)CH.sub.2CH(CH.sub.3).sub.2),
3-methyl-3-pentyl (--C(CH.sub.3)(CH.sub.2CH.sub.3).sub.2),
2-methyl-3-pentyl (--CH(CH.sub.2CH.sub.3)CH(CH.sub.3).sub.2),
2,3-dimethyl-2-butyl (--C(CH.sub.3).sub.2CH(CH.sub.3).sub.2),
3,3-dimethyl-2-butyl (--CH(CH.sub.3)C(CH.sub.3).sub.3, and octyl
(--(CH.sub.2).sub.7CH.sub.3).
[0099] "Alkoxy" means a group having the formula --O-alkyl, in
which an alkyl group, as defined above, is attached to the parent
molecule via an oxygen atom. The alkyl portion of an alkoxy group
can have 1 to 20 carbon atoms (i.e., C.sub.1-C.sub.20 alkoxy), 1 to
12 carbon atoms (i.e., C.sub.1-C.sub.12 alkoxy), or 1 to 6 carbon
atoms (i.e., C.sub.1-C.sub.6 alkoxy). Examples of suitable alkoxy
groups include, but are not limited to, methoxy (--O--CH.sub.3 or
--OMe), ethoxy (--OCH.sub.2CH.sub.3 or --OEt), t-butoxy
(--O--C(CH.sub.3).sub.3 or -OtBu), and the like.
[0100] "Haloalkyl" is an alkyl group, as defined above, in which
one or more hydrogen atoms of the alkyl group is replaced with a
halogen atom. The alkyl portion of a haloalkyl group can have 1 to
20 carbon atoms (i.e., C.sub.1-C.sub.20 haloalkyl), 1 to 12 carbon
atoms (i.e., C.sub.1-C.sub.12 haloalkyl), or 1 to 6 carbon atoms
(i.e., C.sub.1-C.sub.6 alkyl). Examples of suitable haloalkyl
groups include, but are not limited to, --CF.sub.3, --CHF.sub.2,
--CFH.sub.2, --CH.sub.2CF.sub.3, and the like.
[0101] "Alkenyl" is a hydrocarbon containing normal, secondary,
tertiary, or cyclic carbon atoms with at least one site of
unsaturation, i.e. a carbon-carbon, sp2 double bond. For example,
an alkenyl group can have 2 to 20 carbon atoms (i.e.,
C.sub.2-C.sub.20 alkenyl), 2 to 12 carbon atoms (i.e.,
C.sub.2-C.sub.12 alkenyl), or 2 to 6 carbon atoms (i.e.,
C.sub.2-C.sub.6 alkenyl). Examples of suitable alkenyl groups
include, but are not limited to, vinyl (--CH.dbd.CH.sub.2), allyl
(--CH.sub.2CH.dbd.CH.sub.2), cyclopentenyl (--C.sub.5H.sub.7), and
5-hexenyl (--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.dbd.CH.sub.2).
[0102] "Alkynyl" is a hydrocarbon containing normal, secondary,
tertiary or cyclic carbon atoms with at least one site of
unsaturation, i.e. a carbon-carbon, sp triple bond. For example, an
alkynyl group can have 2 to 20 carbon atoms (i.e., C.sub.2-C.sub.20
alkynyl), 2 to 12 carbon atoms (i.e., C.sub.2-C.sub.12 alkyne,), or
2 to 6 carbon atoms (i.e., C.sub.2-C.sub.6 alkynyl). Examples of
suitable alkynyl groups include, but are not limited to, acetylenic
(--C.ident.CH), propargyl (--CH.sub.2C.ident.CH), and the like.
[0103] "Alkylene" refers to a saturated, branched or straight chain
radical or cyclic hydrocarbon radical having two monovalent radical
centers derived by the removal of two hydrogen atoms from the same
or two different carbon atoms of a parent alkane. For example, an
alkylene group can have 1 to 20 carbon atoms, 1 to 10 carbon atoms,
or 1 to 6 carbon atoms. Typical alkylene radicals include, but are
not limited to, methylene (--CH.sub.2--), 1,1-ethylene
(--CH(CH.sub.3)--), 1,2-ethylene (--CH.sub.2CH.sub.2--),
1,1-propylene (--CH(CH.sub.2CH.sub.3)--), 1,2-propylene
(--CH.sub.2CH(CH.sub.3)--), 1,3-propylene
(--CH.sub.2CH.sub.2CH.sub.2--), 1,4-butylene
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--), and the like.
[0104] "Alkenylene" refers to an unsaturated, branched or straight
chain or cyclic hydrocarbon radical having two monovalent radical
centers derived by the removal of two hydrogen atoms from the same
or two different carbon atoms of a parent alkene. For example, and
alkenylene group can have 1 to 20 carbon atoms, 1 to 10 carbon
atoms, or 1 to 6 carbon atoms. Typical alkenylene radicals include,
but are not limited to, 1,2-ethylene (--CH.dbd.CH--).
[0105] "Alkynylene" refers to an unsaturated, branched or straight
chain or cyclic hydrocarbon radical having two monovalent radical
centers derived by the removal of two hydrogen atoms from the same
or two different carbon atoms of a parent alkyne. For example, an
alkynylene group can have 1 to 20 carbon atoms, 1 to 10 carbon
atoms, or 1 to 6 carbon atoms. Typical alkynylene radicals include,
but are not limited to, acetylene (--C.ident.C--), propargyl
(--CH.sub.2C.ident.C--), and 4-pentynyl
(--CH.sub.2CH.sub.2CH.sub.2C.dbd.C--).
[0106] "Alkylyne" refers to a saturated, branched or straight chain
radical having two radical centers derived by the removal of three
hydrogen atoms from two carbon atoms of a parent alkane. For
example, an alkylyne group can have 2 to 20 carbon atoms, 2 to 10
carbon atoms, or 2 to 6 carbon atoms. Typical alkylyne radicals
include, but are not limited to, 1,2-ethylyne (--CH.sub.2CH.dbd.),
1,2-propylyne (--CH.sub.2C(CH.sub.3).dbd.), 1,3-propylyne
(--CH.sub.2CH.sub.2CH.dbd.), 1,4-butylyne
(--CH.sub.2CH.sub.2CH.sub.2CH.dbd.), and the like.
[0107] "Aryl" means a monovalent aromatic hydrocarbon radical
derived by the removal of one hydrogen atom from a single carbon
atom of a parent aromatic ring system. For example, an aryl group
can have 6 to 20 carbon atoms, 6 to 14 carbon atoms, or 6 to 12
carbon atoms. Typical aryl groups include, but are not limited to,
radicals derived from benzene (e.g., phenyl), substituted benzene,
naphthalene, anthracene, biphenyl, and the like.
[0108] "Arylene" refers to an aryl as defined above having two
monovalent radical centers derived by the removal of two hydrogen
atoms from the same or two different carbon atoms of a parent aryl.
Typical arylene radicals include, but are not limited to,
phenylene.
[0109] "Arylalkyl" refers to an acyclic alkyl radical in which one
of the hydrogen atoms bonded to a carbon atom, typically a terminal
or sp3 carbon atom, is replaced with an aryl radical. Typical
arylalkyl groups include, but are not limited to, benzyl,
2-phenylethan-1-yl, naphthylmethyl, 2-naphthylethan-1-yl,
naphthobenzyl, 2-naphthophenylethan-1-yl and the like. The
arylalkyl group can comprise 6 to 20 carbon atoms, e.g., the alkyl
moiety is 1 to 6 carbon atoms and the aryl moiety is 6 to 14 carbon
atoms.
[0110] "Cycloalkyl" refers to a saturated or partially unsaturated
ring having 3 to 7 carbon atoms as a monocycle, 7 to 12 carbon
atoms as a bicycle, and up to about 20 carbon atoms as a polycycle.
Monocyclic cycloalkyl groups have 3 to 6 ring atoms, still more
typically 5 or 6 ring atoms. Bicyclic cycloalkyl groups have 7 to
12 ring atoms, e.g., arranged as a bicyclo (4,5), (5,5), (5,6) or
(6,6) system, or 9 or 10 ring atoms arranged as a bicyclo (5,6) or
(6,6) system. Cycloalkyl groups include hydrocarbon mono-, bi-, and
poly-cyclic rings, whether fused, bridged, or spiro. Non-limiting
examples of monocyclic carbocycles include cyclopropyl, cyclobutyl,
cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl,
1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl,
1-cyclohex-2-enyl, 1-cyclohex-3-enyl, bicyclo[3.1.0]hex-6-yl and
the like.
[0111] "Cycloalkylene" refers to a cycloalkyl as defined above
having two monovalent radical centers derived by the removal of two
hydrogen atoms from the same or two different carbon atoms of a
parent cycloalkyl. Typical cycloalkylene radicals include, but are
not limited to, cyclopropylene, cyclobutylene, cyclopentylene and
cyclohexylene.
[0112] "Cycloalkylyne" refers to a cycloalkyl as defined above
having two radical centers derived by the removal of three hydrogen
atoms from two carbon atoms of a parent cycloalkyl. Two of the
hydrogen atoms are removed from the same carbon atom and one
hydrogen atom is removed from an alternative carbon atom of the
ring. Non-limiting examples of cycloalkylyne radicals include:
##STR00057##
[0113] "Halogen" refers to F, Cl, Br, or I.
[0114] As used herein the term "haloalkyl" refers to an alkyl
group, as defined herein, that is substituted with at least one
halogen. Examples of branched or straight chained "haloalkyl"
groups as used herein include, but are not limited to, methyl,
ethyl, propyl, isopropyl, n-butyl, and t-butyl substituted
independently with one or more halogens, for example, fluoro,
chloro, bromo, and iodo. The term "haloalkyl" should be interpreted
to include such substituents as perfluoroalkyl groups such as
--CF.sub.3.
[0115] As used herein, the term "haloalkoxy" refers to a group
--OR.sup.a, where R.sup.a is a haloalkyl group as herein defined.
As non-limiting examples, haloalkoxy groups include --O(CH.sub.2)F,
--O(CH)F.sub.2, and --OCF.sub.3.
[0116] "Heterocycle" or "heterocyclyl" refers to a saturated or
partially saturated cyclic group having from 1 to 14 carbon atoms
and from 1 to 6 heteroatoms selected from N, S, P, or O, and
includes single ring and multiple ring systems including, fused,
bridged, and spiro ring systems. "Heterocycle" or "heterocyclyl" as
used herein includes by way of example and not limitation those
heterocycles described in Paquette, Leo A.; Principles of Modern
Heterocyclic Chemistry (W. A. Benjamin, New York, 1968),
particularly Chapters 1, 3, 4, 6, 7, and 9; The Chemistry of
Heterocyclic Compounds, A Series of Monographs" (John Wiley &
Sons, New York, 1950 to present), in particular Volumes 13, 14, 16,
19, and 28; and J. Am. Chem. Soc. (1960) 82:5566. In one
embodiment, the carbon, nitrogen, phosphorous, or sulfur atom(s) of
the heterocyclic group may be oxidized to provide for C(.dbd.O),
N-oxide, phosphinane oxide, sulfinyl, or sulfonyl moieties.
[0117] As one example, substituted heterocyclyls include, for
example, heterocyclic rings substituted with any of the
substituents disclosed herein including oxo groups. A non-limiting
example of a carbonyl substituted heterocyclyl is:
##STR00058##
[0118] Examples of heterocycles include by way of example and not
limitation dihydroypyridyl, tetrahydropyridyl(piperidyl),
tetrahydrothiophenyl, sulfur oxidized tetrahydrothiophenyl,
piperidinyl, 4-piperidonyl, pyrrolidinyl, azetidinyl,
2-pyrrolidonyl, tetrahydrofuranyl, decahydroquinolinyl,
octahydroisoquinolinyl, pyranyl, morpholinyl, and
bis-tetrahydrofuranyl:
##STR00059##
[0119] "Heterocyclene" or "heterocyclylene" refers to a
"heterocycle" or "heterocyclyl" as defined above having two
monovalent radical centers derived by the removal of two hydrogen
atoms from the same or two different carbon atoms of a parent
heterocycle, the removal of two hydrogen atoms from two nitrogen
atoms of a parent heterocycle, or the removal of a hydrogen atom
from a nitrogen and the removal of a hydrogen atom from a carbon
atom of a parent heterocycle. Non-limiting examples of
heterocyclene or heterocyclylenes are:
##STR00060##
[0120] "Heterocyclelyne" or "heterocyclylyne" refers to a
"heterocycle" or "heterocyclyl" as defined above having two radical
centers derived by the removal of three hydrogen atoms from two
carbon atoms of a parent heterocycle or the removal of a hydrogen
atom from a nitrogen atom and the removal of two hydrogen atoms
from the same carbon atom of a parent heterocycle. Non-limiting
examples of heterocyclelyne or "heterocyclylyne radicals
include:
##STR00061##
[0121] "Heteroaryl" refers to a monovalent aromatic heterocyclyl
having at least one heteroatom in the ring. Thus, "heteroaryl"
refers to an aromatic group of from 1 to 14 carbon atoms and 1 to 6
heteroatoms selected from oxygen, nitrogen, sulfur, or phosphorous.
For multiple ring systems, by way of example, the term "heteroaryl"
includes fused, bridged, and spiro ring systems having aromatic and
non-aromatic rings. In one embodiment, the carbon, nitrogen, or
sulfur ring atom(s) of the heteroaryl group may be oxidized to
provide for C(.dbd.O), N-oxide, sulfinyl, or sulfonyl moieties.
[0122] Examples of heteroaryls include by way of example and not
limitation pyridyl, thiazolyl, pyrimidinyl, furanyl, thienyl,
pyrrolyl, pyrazolyl, imidazolyl, tetrazolyl, benzofuranyl,
thianaphthalenyl, indolyl, quinolinyl, isoquinolinyl,
benzimidazolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl,
6H-1,2,5-thiadiazinyl, 2H,6H-1,5,2-dithiazinyl, thienyl,
thianthrenyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxathinyl,
2H-pyrrolyl, isothiazolyl, isoxazolyl, pyrazinyl, pyridazinyl,
indolizinyl, isoindolyl, 3H-indolyl, 1H-indazoly, purinyl,
4H-quinolizinyl, naphthyridinyl, quinoxalinyl, quinazolinyl,
cinnolinyl, pteridinyl, 4aH-carbazolyl, carbazolyl,
.beta.-carbolinyl, phenanthridinyl, acridinyl, pyrimidinyl,
phenanthrolinyl, phenazinyl, phenothiazinyl, furazanyl,
phenoxazinyl, isochromanyl, chromanyl, imidazolidinyl,
imidazolinyl, pyrazolidinyl, pyrazolinyl, piperazinyl, indolinyl,
isoindolinyl, quinuclidinyl, morpholinyl, oxazolidinyl,
benzotriazolyl, benzisoxazolyl, oxindolyl, benzoxazolinyl, and
isatinoyl. "Heterocyclylene" refers to a heterocyclyl, as defined
herein, derived by replacing a hydrogen atom from a carbon atom or
heteroatom of a heterocyclyl, with an open valence. Similarly,
"heteroarylene" refers to an aromatic heterocyclylene.
[0123] "Heterocyclylalkyl" refers to an acyclic alkyl radical in
which one of the hydrogen atoms bonded to a carbon atom, typically
a terminal or sp3 carbon atom, is replaced with a heterocyclyl
radical (i.e., a heterocyclyl-alkylene-moiety). Typical
heterocyclyl alkyl groups include, but are not limited to
heterocyclyl-CH.sub.2--, 2-(heterocyclyl)ethan-1-yl, and the like,
wherein the "heterocyclyl" portion includes any of the heterocyclyl
groups described above, including those described in Principles of
Modern Heterocyclic Chemistry. One skilled in the art will also
understand that the heterocyclyl group can be attached to the alkyl
portion of the heterocyclyl alkyl by means of a carbon-carbon bond
or a carbon-heteroatom bond, with the proviso that the resulting
group is chemically stable. The heterocyclylalkyl group comprises 2
to 20 carbon atoms and 1-6 heteroatoms, e.g., the alkyl portion of
the heterocyclylalkyl group comprises 1 to 6 carbon atoms and the
heterocyclyl moiety comprises 1 to 14 carbon atoms. Examples of
heterocyclylalkyls include by way of example and not limitation
5-membered sulfur, oxygen, phosphorus, and/or nitrogen containing
heterocycles such as pyrrolidiylmethyl,
2-tetrahydrofuranylylethan-1-yl, and the like, 6-membered sulfur,
oxygen, and/or nitrogen containing heterocycles such as
piperidinylmethyl, morpholinylmethyl, piperidinylethyl,
teterahydropyranylethyl, and the like.
[0124] "Heteroarylalkyl" refers to an alkyl group, as defined
herein, in which a hydrogen atom has been replaced with a
heteroaryl group as defined herein. Non-limiting examples of
heteroaryl alkyl include --CH.sub.2-pyridinyl, --CH.sub.2-pyrrolyl,
--CH.sub.2-oxazolyl, --CH.sub.2-indolyl, --CH.sub.2-isoindolyl,
--CH.sub.2-purinyl, --CH.sub.2-furanyl, --CH.sub.2-thienyl,
--CH.sub.2-benzofuranyl, --CH.sub.2-benzothiophenyl,
--CH.sub.2-carbazolyl, --CH.sub.2-imidazolyl, --CH.sub.2-thiazolyl,
--CH.sub.2-isoxazolyl, --CH.sub.2-pyrazolyl,
--CH.sub.2-isothiazolyl, --CH.sub.2-quinolyl,
--CH.sub.2-isoquinolyl, --CH.sub.2-pyridazyl, --CH.sub.2-pyrimidyl,
--CH.sub.2-pyrazyl, --CH(CH.sub.3)-pyridinyl,
--CH(CH.sub.3)-pyrrolyl, --CH(CH.sub.3)-oxazolyl,
--CH(CH.sub.3)-indolyl, --CH(CH.sub.3)-isoindolyl,
--CH(CH.sub.3)-purinyl, --CH(CH.sub.3)-furanyl,
--CH(CH.sub.3)-thienyl, --CH(CH.sub.3)-benzofuranyl,
--CH(CH.sub.3)-benzothiophenyl, --CH(CH.sub.3)-carbazolyl,
--CH(CH.sub.3)-imidazolyl, --CH(CH.sub.3)-thiazolyl,
--CH(CH.sub.3)-isoxazolyl, --CH(CH.sub.3)-pyrazolyl,
--CH(CH.sub.3)-isothiazolyl, --CH(CH.sub.3)-quinolyl,
--CH(CH.sub.3)-isoquinolyl, --CH(CH.sub.3)-pyridazyl,
--CH(CH.sub.3)-pyrimidyl, --CH(CH.sub.3)-pyrazyl, and the like.
[0125] The term "heterocyclyloxy" represents a heterocyclyl group
attached to the adjacent atom by an oxygen.
[0126] When there is a sulfur atom present, the sulfur atom can be
at different oxidation levels, namely, S, SO, SO.sub.2, or
SO.sub.3. All such oxidation levels are within the scope of the
present invention.
[0127] When there is a phosphorous atom present, the phosphorous
atom can be at different oxidation levels, namely,
POR.sup.aR.sup.bR.sup.b, PO.sub.2R.sup.aR.sup.b, or
PO.sub.3R.sup.aR.sup.b, where R.sup.a, R.sup.b, and R.sup.c each
independently is chosen from H, C.sub.1-12 alkyl, C.sub.2-12
alkenyl, C.sub.2-12 alkynyl, C.sub.6-14 aryl, 3-12 membered
heterocycle, 3-18 membered heteroarylalkyl, C.sub.6-18 arylalkyl;
or two taken together (with or without oxygens) form a 5 to 10
membered heterocycle. All such oxidation levels are within the
scope of the present invention
[0128] The term "optionally substituted" in reference to a
particular moiety of the compound of the Formulae of the invention,
for example an "optionally substituted aryl group", refers to a
moiety having none, one, or more substituents.
[0129] The term "substituted" in reference to alkyl, alkylene,
aryl, arylalkyl, alkoxy, heterocyclyl, heteroaryl, carbocyclyl,
etc., for example, "substituted alkyl", "substituted alkylene",
"substituted aryl", "substituted arylalkyl", "substituted
heterocyclyl", and "substituted carbocyclyl" means alkyl, alkylene,
aryl, arylalkyl, heterocyclyl, carbocyclyl respectively, in which
one or more hydrogen atoms are each independently replaced with a
non-hydrogen substituent. Divalent groups may also be similarly
substituted. Unless otherwise indicated, typical substituents
include, but are not limited to, --X, --R.sup.b, --O.sup.-, .dbd.O,
--OR.sup.b, --SR.sup.b, --S.sup.-, --NR.sup.b.sub.2,
--N.sup.+R.sup.b.sub.3, .dbd.NR.sup.b, --CX.sub.3, --CN, --OCN,
--SCN, --N.dbd.C.dbd.O, --NCS, --NO, --NO.sub.2, .dbd.N.sub.2,
--N.sub.3, --NHC(.dbd.O)R.sup.b, --OC(.dbd.O)R.sup.b,
--NHC(.dbd.O)NR.sup.b.sub.2, --S(.dbd.O).sub.2--,
--S(.dbd.O).sub.2OH, --S(.dbd.O).sub.2R.sup.b,
--OS(.dbd.O).sub.2OR.sup.b, --S(.dbd.O).sub.2NR.sup.b.sub.2,
--S(.dbd.O)R.sup.b, --OP(.dbd.O)(OR.sup.b).sub.2,
--P(.dbd.O)(OR.sup.b).sub.2, --P(.dbd.O)(O.sup.-).sub.2,
--P(.dbd.O)(OH).sub.2, --P(O)(OR.sup.b)(O.sup.-),
--C(.dbd.O)R.sup.b, --C(.dbd.O)X, --C(S)R.sup.b, --C(O)OR.sup.b,
--C(O)O.sup.-, --C(S)OR.sup.b, --C(O)SR.sup.b, --C(S)SR.sup.b,
--C(O)NR.sup.b.sub.2, --C(S)NR.sup.b.sub.2,
--C(.dbd.NR.sup.b)NR.sup.b.sub.2, where each X is independently a
halogen: F, Cl, Br, or I; and each R.sup.b is independently H,
alkyl, aryl, arylalkyl, a heterocycle, or a protecting group or
prodrug moiety. Alkylene, alkenylene, and alkynylene groups may
also be similarly substituted. Unless otherwise indicated, when the
term "substituted" is used in conjunction with groups such as
arylalkyl, which have two or more moieties capable of substitution,
the substituents can be attached to the aryl moiety, the alkyl
moiety, or both.
[0130] Those skilled in the art will recognize that when moieties
such as "alkyl", "aryl", "heterocyclyl", etc. are substituted with
one or more substituents, they could alternatively be referred to
as "alkylene", "arylene", "heterocyclylene", etc. moieties (i.e.,
indicating that at least one of the hydrogen atoms of the parent
"alkyl", "aryl", "heterocyclyl" moieties has been replaced with the
indicated substituent(s)). When moieties such as "alkyl", "aryl",
"heterocyclyl", etc. are referred to herein as "substituted" or are
shown diagrammatically to be substituted (or optionally
substituted, e.g., when the number of substituents ranges from zero
to a positive integer), then the terms "alkyl", "aryl",
"heterocyclyl", etc. are understood to be interchangeable with
"alkylene", "arylene", "heterocyclylene", etc.
[0131] As will be appreciated by those skilled in the art, the
compounds of the present invention may exist in solvated or
hydrated form. The scope of the present invention includes such
forms. Again, as will be appreciated by those skilled in the art,
the compounds may be capable of esterification. The scope of the
present invention includes esters and other physiologically
functional derivatives. The scope of the present invention includes
prodrug forms of the compound herein described.
[0132] "Ester" means any ester of a compound in which any of the
--COON functions of the molecule is replaced by a --C(O)OR
function, or in which any of the --OH functions of the molecule are
replaced with a --OC(O)R function, in which the R moiety of the
ester is any carbon-containing group which forms a stable ester
moiety, including but not limited to alkyl, alkenyl, alkynyl,
cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclyl,
heterocyclylalkyl and substituted derivatives thereof.
[0133] The term "prodrug" as used herein refers to any compound
that when administered to a biological system generates the drug
substance, i.e., active ingredient, as a result of spontaneous
chemical reaction(s), enzyme catalyzed chemical reaction(s),
photolysis, and/or metabolic chemical reaction(s). A prodrug is
thus a covalently modified analog or latent form of a
therapeutically active compound. Example of prodrugs include ester
moieties, quaternary ammonium moieties, glycol moieties, and the
like.
[0134] One skilled in the art will recognize that substituents and
other moieties of the compounds of Formula I should be selected in
order to provide a compound which is sufficiently stable to provide
a pharmaceutically useful compound which can be formulated into an
acceptably stable pharmaceutical composition. Compounds of Formula
I which have such stability are contemplated as falling within the
scope of the present invention.
[0135] As will be appreciated by those skilled in the art, the
compounds of the present invention may contain one or more chiral
centers. The scope of the present invention includes such forms.
Again, as will be appreciated by those skilled in the art, the
compound is capable of esterification. The scope of the present
invention includes esters and other physiologically functional
derivatives. In addition, the scope of the present invention
includes prodrug forms of the compound herein described.
[0136] A compound of Formula I-II and its pharmaceutically
acceptable salts may exist as different polymorphs or
pseudopolymorphs. As used herein, crystalline polymorphism means
the ability of a crystalline compound to exist in different crystal
structures. Polymorphism generally can occur as a response to
changes in temperature, pressure, or both. Polymorphism can also
result from variations in the crystallization process. Polymorphs
can be distinguished by various physical characteristics known in
the art such as x-ray diffraction patterns, solubility, and melting
point. The crystalline polymorphism may result from differences in
crystal packing (packing polymorphism) or differences in packing
between different conformers of the same molecule (conformational
polymorphism). As used herein, crystalline pseudopolymorphism means
the ability of a hydrate or solvate of a compound to exist in
different crystal structures. The pseudopolymorphs of the instant
invention may exist due to differences in crystal packing (packing
pseudopolymorphism) or due to differences in packing between
different conformers of the same molecule (conformational
pseudopolymorphism). The instant invention comprises all polymorphs
and pseudopolymorphs of the compounds of Formula I-II and their
pharmaceutically acceptable salts.
[0137] A compound of Formula I-II and its pharmaceutically
acceptable salts may also exist as an amorphous solid. As used
herein, an amorphous solid is a solid in which there is no
long-range order of the positions of the atoms in the solid. This
definition applies as well when the crystal size is two nanometers
or less. Additives, including solvents, may be used to create the
amorphous forms of the instant invention. The instant invention
comprises all amorphous forms of the compounds of Formula I-II and
their pharmaceutically acceptable salts.
[0138] Certain of the compounds described herein contain one or
more chiral centers, or may otherwise be capable of existing as
multiple stereoisomers. The scope of the present invention includes
mixtures of stereoisomers as well as purified enantiomers or
enantiomerically/diastereomerically enriched mixtures. Also
included within the scope of the invention are the individual
isomers of the compounds represented by the formulae of the present
invention, as well as any wholly or partially equilibrated mixtures
thereof. The present invention also includes the individual isomers
of the compounds represented by the formulas above as mixtures with
isomers thereof in which one or more chiral centers are
inverted.
[0139] The term "chiral" refers to molecules which have the
property of non-superimposability of the mirror image partner,
while the term "achiral" refers to molecules which are
superimposable on their mirror image partner.
[0140] The term "stereoisomers" refers to compounds which have
identical chemical constitution, but differ with regard to the
arrangement of the atoms or groups in space.
[0141] "Diastereomer" refers to a stereoisomer with two or more
centers of chirality and whose molecules are not mirror images of
one another. Diastereomers have different physical properties,
e.g., melting points, boiling points, spectral properties, and
reactivities. Mixtures of diastereomers may separate under high
resolution analytical procedures such as electrophoresis and
chromatography.
[0142] "Enantiomers" refer to stereoisomers of a compound which are
non-superimposable mirror images of one another.
[0143] "Atropisomers" refer to stereoisomers of a compound
resulting from hindered rotation about single bonds where the
steric strain barrier to rotation is high enough to allow for the
isolation of the individual conformer. Atropisomers display axial
chirality. Atropisomers may be equilibrated thermally and the
interconversion barrier may be measured kinetically. Atropisomerism
may occur apart from the presence of other forms of chiral
isomerism. Thus, as illustrated, the depicted nitrogen atom is
planar and compounds of Formula I are capable of existing as
atropisomers:
##STR00062##
[0144] In one embodiment of the present invention, the compounds
exist in a conformeric form of Formula Ia:
##STR00063##
[0145] Stereochemical definitions and conventions used herein
generally follow S. P. Parker, Ed., McGraw-Hill Dictionary of
Chemical Terms (1984) McGraw-Hill Book Company, New York; and
Eliel, E. and Wilen, S., Stereochemistry of Organic Compounds
(1994) John Wiley & Sons, Inc., New York.
[0146] Many organic compounds exist in optically active forms,
i.e., they have the ability to rotate the plane of plane-polarized
light. In describing an optically active compound, the prefixes D
and L or R and S are used to denote the absolute configuration of
the molecule about its chiral center(s). The prefixes d and l or
(+) and (-) are employed to designate the sign of rotation of
plane-polarized light by the compound, with (-) or 1 meaning that
the compound is levorotatory. A compound prefixed with (+) or d is
dextrorotatory.
[0147] A specific stereoisomer may also be referred to as an
enantiomer, and a mixture of such isomers is often called an
enantiomeric mixture. A 50:50 mixture of enantiomers is referred to
as a racemic mixture or a racemate, which may occur where there has
been no stereoselection or stereospecificity in a chemical reaction
or process. The terms "racemic mixture" and "racemate" refer to an
equimolar mixture of two enantiomeric species, devoid of optical
activity.
[0148] The present invention includes a salt or solvate of the
compounds herein described, including combinations thereof such as
a solvate of a salt. The compounds of the present invention may
exist in solvated, for example hydrated, as well as unsolvated
forms, and the present invention encompasses all such forms.
[0149] Typically, but not absolutely, the salts of the present
invention are pharmaceutically acceptable salts. Salts encompassed
within the term "pharmaceutically acceptable salts" refer to
non-toxic salts of the compounds of this invention.
[0150] Examples of suitable pharmaceutically acceptable salts
include inorganic acid addition salts such as chloride, bromide,
sulfate, phosphate, and nitrate; organic acid addition salts such
as acetate, galactarate, propionate, succinate, lactate, glycolate,
malate, tartrate, citrate, maleate, fumarate, methanesulfonate,
p-toluenesulfonate, and ascorbate; salts with acidic amino acid
such as aspartate and glutamate; alkali metal salts such as sodium
salt and potassium salt; alkaline earth metal salts such as
magnesium salt and calcium salt; ammonium salt; organic basic salts
such as trimethylamine salt, triethylamine salt, pyridine salt,
picoline salt, dicyclohexylamine salt, and
N,N'-dibenzylethylenediamine salt; and salts with basic amino acid
such as lysine salt and arginine salt. The salts may be in some
cases hydrates or ethanol solvates.
[0151] The modifier "about" used in connection with a quantity is
inclusive of the stated value and has the meaning dictated by the
context (e.g., includes the degree of error associated with
measurement of the particular quantity).
[0152] Whenever a compound described herein is substituted with
more than one of the same designated group, e.g., "R" or "R.sup.1",
then it will be understood that the groups may be the same or
different, i.e., each group is independently selected. Wavy
lines,
##STR00064##
indicate the site of covalent bond attachments to the adjoining
substructures, groups, moieties, or atoms.
[0153] The compounds of the invention can also exist as tautomeric
isomers in certain cases. Although only one delocalized resonance
structure may be depicted, all such forms are contemplated within
the scope of the invention. For example, ene-amine tautomers can
exist for purine, pyrimidine, imidazole, guanidine, amidine, and
tetrazole systems and all their possible tautomeric forms are
within the scope of the invention.
[0154] Selected substituents comprising the compounds of Formula
I-II may be present to a recursive degree. In this context,
"recursive substituent" means that a substituent may recite another
instance of itself. The multiple recitations may be direct or
indirect through a sequence of other substituents. Because of the
recursive nature of such substituents, theoretically, a large
number of compounds may be present in any given embodiment. One of
ordinary skill in the art of medicinal chemistry understands that
the total number of such substituents is reasonably limited by the
desired properties of the compound intended. Such properties
include, by way of example and not limitation, physical properties
such as molecular weight, solubility or log P, application
properties such as activity against the intended target, and
practical properties such as ease of synthesis. Recursive
substituents may be an intended aspect of the invention. One of
ordinary skill in the art of medicinal chemistry understands the
versatility of such substituents. To the degree that recursive
substituents are present in an embodiment of the invention, they
may recite another instance of themselves, 0, 1, 2, 3, or 4
times.
[0155] The compounds of Formula I-II also include molecules that
incorporate isotopes of the atoms specified in the particular
molecules. Non-limiting examples of these isotopes include D, T,
.sup.14C, .sup.13C, .sup.18O and .sup.15N.
Protecting Groups
[0156] In the context of the present invention, protecting groups
include prodrug moieties and chemical protecting groups.
[0157] Protecting groups are available, commonly known and used,
and are optionally used to prevent side reactions with the
protected group during synthetic procedures, i.e. routes or methods
to prepare the compounds of the invention. For the most part the
decision as to which groups to protect, when to do so, and the
nature of the chemical protecting group "PG" will be dependent upon
the chemistry of the reaction to be protected against (e.g.,
acidic, basic, oxidative, reductive or other conditions) and the
intended direction of the synthesis. The PG groups do not need to
be, and generally are not, the same if the compound is substituted
with multiple PG. In general, PG will be used to protect functional
groups such as carboxyl, hydroxyl, thio, or amino groups and to
thus prevent side reactions or to otherwise facilitate the
synthetic efficiency. The order of deprotection to yield free,
deprotected groups is dependent upon the intended direction of the
synthesis and the reaction conditions to be encountered, and may
occur in any order as determined by the artisan.
[0158] Various functional groups of the compounds of the invention
may be protected. For example, protecting groups for --OH groups
(whether hydroxyl, carboxylic acid, phosphonic acid, or other
functions) include "ether- or ester-forming groups". Ether- or
ester-forming groups are capable of functioning as chemical
protecting groups in the synthetic schemes set forth herein.
However, some hydroxyl and thio protecting groups are neither
ether-nor ester-forming groups, as will be understood by those
skilled in the art, and are included with amides, discussed
below.
[0159] A very large number of hydroxyl protecting groups and
amide-forming groups and corresponding chemical cleavage reactions
are described in Protective Groups in Organic Synthesis, Theodora
W. Greene and Peter G. M. Wuts (John Wiley & Sons, Inc., New
York, 1999, ISBN 0-471-16019-9) ("Greene"). See also Kocienski,
Philip J.; Protecting Groups (Georg Thieme Verlag Stuttgart, New
York, 1994), which is incorporated by reference in its entirety
herein. In particular Chapter 1, Protecting Groups: An Overview,
pages 1-20, Chapter 2, Hydroxyl Protecting Groups, pages 21-94,
Chapter 3, Diol Protecting Groups, pages 95-117, Chapter 4,
Carboxyl Protecting Groups, pages 118-154, Chapter 5, Carbonyl
Protecting Groups, pages 155-184. For protecting groups for
carboxylic acid, phosphonic acid, phosphonate, sulfonic acid and
other protecting groups for acids see Greene as set forth below.
Such groups include by way of example and not limitation, esters,
amides, hydrazides, and the like.
Ether- and Ester-Forming Protecting Groups
[0160] Ester-forming groups include: (1) phosphonate ester-forming
groups, such as phosphonamidate esters, phosphorothioate esters,
phosphonate esters, and phosphon-bis-amidates; (2) carboxyl
ester-forming groups, and (3) sulphur ester-forming groups, such as
sulphonate, sulfate, and sulfinate.
Metabolites of the Compounds of the Invention
[0161] Also falling within the scope of this invention are the in
vivo metabolic products of the compounds described herein. Such
products may result for example from the oxidation, reduction,
hydrolysis, amidation, esterification and the like of the
administered compound, primarily due to enzymatic processes.
Accordingly, the invention includes compounds produced by a process
comprising contacting a compound of this invention with a mammal
for a period of time sufficient to yield a metabolic product
thereof. Such products typically are identified by preparing a
radiolabelled (e.g., C.sup.14 or .sup.3H) compound of the
invention, administering it parenterally in a detectable dose
(e.g., greater than about 0.5 mg/kg) to an animal such as rat,
mouse, guinea pig, monkey, or to man, allowing sufficient time for
metabolism to occur (typically about 30 seconds to 30 hours) and
isolating its conversion products from the urine, blood or other
biological samples. These products are easily isolated since they
are labeled (others are isolated by the use of antibodies capable
of binding epitopes surviving in the metabolite). The metabolite
structures are determined in conventional fashion, e.g., by MS or
NMR analysis. In general, analysis of metabolites is done in the
same way as conventional drug metabolism studies well-known to
those skilled in the art. The conversion products, so long as they
are not otherwise found in vivo, are useful in diagnostic assays
for therapeutic dosing of the compounds of the invention even if
they possess no anti-infective activity of their own.
[0162] The definitions and substituents for various genus and
subgenus of the present compounds are described and illustrated
herein. It should be understood by one skilled in the art that any
combination of the definitions and substituents described above
should not result in an inoperable species or compound. "Inoperable
species or compounds" means compound structures that violates
relevant scientific principles (such as, for example, a carbon atom
connecting to more than four covalent bonds) or compounds too
unstable to permit isolation and formulation into pharmaceutically
acceptable dosage forms.
Pharmaceutical Formulations
[0163] The compounds of this invention are formulated with
conventional carriers and excipients, which will be selected in
accord with ordinary practice. Tablets will contain excipients,
glidants, fillers, binders and the like. Aqueous formulations are
prepared in sterile form, and when intended for delivery by other
than oral administration generally will be isotonic. All
formulations will optionally contain excipients such as those set
forth in the Handbook of Pharmaceutical Excipients (1986), herein
incorporated by reference in its entirety. Excipients include
ascorbic acid and other antioxidants, chelating agents such as
EDTA, carbohydrates such as dextrin, hydroxyalkylcellulose,
hydroxyalkylmethylcellulose, stearic acid and the like. The pH of
the formulations ranges from about 3 to about 11, but is ordinarily
about 7 to 10.
[0164] While it is possible for the active ingredients to be
administered alone it may be preferable to present them as
pharmaceutical formulations. The formulations of the invention,
both for veterinary and for human use, comprise at least one active
ingredient, together with one or more acceptable carriers and
optionally other therapeutic ingredients. The carrier(s) must be
"acceptable" in the sense of being compatible with the other
ingredients of the formulation and physiologically innocuous to the
recipient thereof.
[0165] The formulations include those suitable for the foregoing
administration routes. The formulations may conveniently be
presented in unit dosage form and may be prepared by any of the
methods well known in the art of pharmacy. Techniques and
formulations generally are found in Remington's Pharmaceutical
Sciences (Mack Publishing Co., Easton, Pa.), herein incorporated by
reference in its entirety. Such methods include the step of
bringing into association the active ingredient with the carrier
which constitutes one or more accessory ingredients. In general the
formulations are prepared by uniformly and intimately bringing into
association the active ingredient with liquid carriers or finely
divided solid carriers or both, and then, if necessary, shaping the
product.
[0166] Formulations of the present invention suitable for oral
administration may be presented as discrete units such as capsules,
cachets or tablets each containing a predetermined amount of the
active ingredient; as a powder or granules; as a solution or a
suspension in an aqueous or non-aqueous liquid; or as an
oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The
active ingredient may also be administered as a bolus, electuary or
paste.
[0167] A tablet is made by compression or molding, optionally with
one or more accessory ingredients. Compressed tablets may be
prepared by compressing in a suitable machine the active ingredient
in a free-flowing form such as a powder or granules, optionally
mixed with a binder, lubricant, inert diluent, preservative,
surface active or dispersing agent. Molded tablets may be made by
molding in a suitable machine a mixture of the powdered active
ingredient moistened with an inert liquid diluent. The tablets may
optionally be coated or scored and optionally are formulated so as
to provide slow or controlled release of the active ingredient.
[0168] For administration to the eye or other external tissues
e.g., mouth and skin, the formulations are preferably applied as a
topical ointment or cream containing the active ingredient(s) in an
amount of, for example, 0.075 to 20% w/w (including active
ingredient(s) in a range between 0.1% and 20% in increments of 0.1%
w/w such as 0.6% w/w, 0.7% w/w, etc.), preferably 0.2 to 15% w/w
and most preferably 0.5 to 10% w/w. When formulated in an ointment,
the active ingredients may be employed with either a paraffinic or
a water-miscible ointment base. Alternatively, the active
ingredients may be formulated in a cream with an oil-in-water cream
base.
[0169] If desired, the aqueous phase of the cream base may include,
for example, at least 30% w/w of a polyhydric alcohol, i.e. an
alcohol having two or more hydroxyl groups such as propylene
glycol, butane 1,3-diol, mannitol, sorbitol, glycerol and
polyethylene glycol (including PEG 400) and mixtures thereof. The
topical formulations may desirably include a compound which
enhances absorption or penetration of the active ingredient through
the skin or other affected areas. Examples of such dermal
penetration enhancers include dimethyl sulphoxide and related
analogs.
[0170] The oily phase of the emulsions of this invention may be
constituted from known ingredients in a known manner. While the
phase may comprise merely an emulsifier (otherwise known as an
emulgent), it desirably comprises a mixture of at least one
emulsifier with a fat or an oil or with both a fat and an oil.
Preferably, a hydrophilic emulsifier is included together with a
lipophilic emulsifier which acts as a stabilizer. It is also
preferred to include both an oil and a fat. Together, the
emulsifier(s) with or without stabilizer(s) make up the so-called
emulsifying wax, and the wax together with the oil and fat make up
the so-called emulsifying ointment base which forms the oily
dispersed phase of the cream formulations.
[0171] Emulgents and emulsion stabilizers suitable for use in the
formulation of the invention include Tween.RTM. 60, Span.RTM. 80,
cetostearyl alcohol, benzyl alcohol, myristyl alcohol, glyceryl
mono-stearate and sodium lauryl sulfate.
[0172] The choice of suitable oils or fats for the formulation is
based on achieving the desired cosmetic properties. The cream
should preferably be a non-greasy, non-staining and washable
product with suitable consistency to avoid leakage from tubes or
other containers. Straight or branched chain, mono- or dibasic
alkyl esters such as di-isoadipate, isocetyl stearate, propylene
glycol diester of coconut fatty acids, isopropyl myristate, decyl
oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate
or a blend of branched chain esters known as Crodamol CAP may be
used, the last three being preferred esters. These may be used
alone or in combination depending on the properties required.
Alternatively, high melting point lipids such as white soft
paraffin and/or liquid paraffin or other mineral oils are used.
[0173] Pharmaceutical formulations according to the present
invention comprise one or more compounds of the invention together
with one or more pharmaceutically acceptable carriers or excipients
and optionally other therapeutic agents. Pharmaceutical
formulations containing the active ingredient may be in any form
suitable for the intended method of administration. When used for
oral use for example, tablets, troches, lozenges, aqueous or oil
suspensions, dispersible powders or granules, emulsions, hard or
soft capsules, syrups or elixirs may be prepared. Compositions
intended for oral use may be prepared according to any method known
to the art for the manufacture of pharmaceutical compositions and
such compositions may contain one or more agents including
sweetening agents, flavoring agents, coloring agents and preserving
agents, in order to provide a palatable preparation. Tablets
containing the active ingredient in admixture with non-toxic
pharmaceutically acceptable excipient which are suitable for
manufacture of tablets are acceptable. These excipients may be, for
example, inert diluents, such as calcium or sodium carbonate,
lactose, lactose monohydrate, croscarmellose sodium, povidone,
calcium or sodium phosphate; granulating and disintegrating agents,
such as maize starch, or alginic acid; binding agents, such as
cellulose, microcrystalline cellulose, starch, gelatin or acacia;
and lubricating agents, such as magnesium stearate, stearic acid or
talc. Tablets may be uncoated or may be coated by known techniques
including microencapsulation to delay disintegration and adsorption
in the gastrointestinal tract and thereby provide a sustained
action over a longer period. For example, a time delay material
such as glyceryl monostearate or glyceryl distearate alone or with
a wax may be employed.
[0174] Formulations for oral use may be also presented as hard
gelatin capsules where the active ingredient is mixed with an inert
solid diluent, for example calcium phosphate or kaolin, or as soft
gelatin capsules wherein the active ingredient is mixed with water
or an oil medium, such as peanut oil, liquid paraffin or olive
oil.
[0175] Aqueous suspensions of the invention contain the active
materials in admixture with excipients suitable for the manufacture
of aqueous suspensions. Such excipients include a suspending agent,
such as sodium carboxymethylcellulose, methylcellulose,
hydroxypropyl methylcelluose, sodium alginate,
polyvinylpyrrolidone, gum tragacanth and gum acacia, and dispersing
or wetting agents such as a naturally occurring phosphatide (e.g.,
lecithin), a condensation product of an alkylene oxide with a fatty
acid (e.g., polyoxyethylene stearate), a condensation product of
ethylene oxide with a long chain aliphatic alcohol (e.g.,
heptadecaethyleneoxycetanol), a condensation product of ethylene
oxide with a partial ester derived from a fatty acid and a hexitol
anhydride (e.g., polyoxyethylene sorbitan monooleate). The aqueous
suspension may also contain one or more preservatives such as ethyl
or n-propyl p-hydroxy-benzoate, one or more coloring agents, one or
more flavoring agents and one or more sweetening agents, such as
sucrose or saccharin.
[0176] Oil suspensions may be formulated by suspending the active
ingredient in a vegetable oil, such as arachis oil, olive oil,
sesame oil or coconut oil, or in a mineral oil such as liquid
paraffin. The oral suspensions may contain a thickening agent, such
as beeswax, hard paraffin or cetyl alcohol. Sweetening agents, such
as those set forth herein, and flavoring agents may be added to
provide a palatable oral preparation. These compositions may be
preserved by the addition of an antioxidant such as ascorbic
acid.
[0177] Dispersible powders and granules of the invention suitable
for preparation of an aqueous suspension by the addition of water
provide the active ingredient in admixture with a dispersing or
wetting agent, a suspending agent, and one or more preservatives.
Suitable dispersing or wetting agents and suspending agents are
exemplified by those disclosed above. Additional excipients, for
example sweetening, flavoring and coloring agents, may also be
present.
[0178] The pharmaceutical compositions of the invention may also be
in the form of oil-in-water emulsions. The oily phase may be a
vegetable oil, such as olive oil or arachis oil, a mineral oil,
such as liquid paraffin, or a mixture of these. Suitable
emulsifying agents include naturally-occurring gums, such as gum
acacia and gum tragacanth, naturally occurring phosphatides, such
as soybean lecithin, esters or partial esters derived from fatty
acids and hexitol anhydrides, such as sorbitan monooleate, and
condensation products of these partial esters with ethylene oxide,
such as polyoxyethylene sorbitan monooleate. The emulsion may also
contain sweetening and flavoring agents. Syrups and elixirs may be
formulated with sweetening agents, such as glycerol, sorbitol or
sucrose. Such formulations may also contain a demulcent, a
preservative, a flavoring or a coloring agent.
[0179] The pharmaceutical compositions of the invention may be in
the form of a sterile injectable preparation, such as a sterile
injectable aqueous or oleaginous suspension. This suspension may be
formulated according to the known art using those suitable
dispersing or wetting agents and suspending agents which have been
mentioned herein. The sterile injectable preparation may also be a
sterile injectable solution or suspension in a non-toxic
parenterally acceptable diluent or solvent, such as a solution in
1,3-butane-diol or prepared as a lyophilized powder. Among the
acceptable vehicles and solvents that may be employed are water,
Ringer's solution and isotonic sodium chloride solution. In
addition, sterile fixed oils may conventionally be employed as a
solvent or suspending medium. For this purpose any bland fixed oil
may be employed including synthetic mono- or diglycerides. In
addition, fatty acids such as oleic acid may likewise be used in
the preparation of injectables.
[0180] The amount of active ingredient that may be combined with
the carrier material to produce a single dosage form will vary
depending upon the host treated and the particular mode of
administration. For example, a time-release formulation intended
for oral administration to humans may contain approximately 1 to
1000 mg of active material compounded with an appropriate and
convenient amount of carrier material which may vary from about 5
to about 95% of the total compositions (weight:weight). The
pharmaceutical composition can be prepared to provide easily
measurable amounts for administration. For example, an aqueous
solution intended for intravenous infusion may contain from about 3
to 500 .mu.g of the active ingredient per milliliter of solution in
order that infusion of a suitable volume at a rate of about 30
mL/hr can occur.
[0181] Formulations suitable for administration to the eye include
eye drops wherein the active ingredient is dissolved or suspended
in a suitable carrier, especially an aqueous solvent for the active
ingredient. The active ingredient is preferably present in such
formulations in a concentration of 0.5 to 20%, advantageously 0.5
to 10% particularly about 1.5% w/w.
[0182] Formulations suitable for topical administration in the
mouth include lozenges comprising the active ingredient in a
flavored basis, usually sucrose and acacia or tragacanth; pastilles
comprising the active ingredient in an inert basis such as gelatin
and glycerin, or sucrose and acacia; and mouthwashes comprising the
active ingredient in a suitable liquid carrier.
[0183] Formulations for rectal administration may be presented as a
suppository with a suitable base comprising for example cocoa
butter or a salicylate.
[0184] Formulations suitable for intrapulmonary or nasal
administration have a particle size for example in the range of 0.1
to 500 .mu.m (including particle sizes in a range between 0.1 and
500 .mu.m in increments such as 0.5 .mu.m, 1 .mu.m, 30 .mu.m, 35
.mu.m, etc.), which is administered by rapid inhalation through the
nasal passage or by inhalation through the mouth so as to reach the
alveolar sacs. Suitable formulations include aqueous or oily
solutions of the active ingredient. Formulations suitable for
aerosol or dry powder administration may be prepared according to
conventional methods and may be delivered with other therapeutic
agents such as compounds heretofore used in the treatment or
prophylaxis of infections as described herein.
[0185] Formulations suitable for vaginal administration may be
presented as pessaries, tampons, creams, gels, pastes, foams or
spray formulations containing in addition to the active ingredient
such carriers as are known in the art to be appropriate.
[0186] Formulations suitable for parenteral administration include
aqueous and non-aqueous sterile injection solutions which may
contain anti-oxidants, buffers, bacteriostats and solutes which
render the formulation isotonic with the blood of the intended
recipient; and aqueous and non-aqueous sterile suspensions which
may include suspending agents and thickening agents.
[0187] The formulations are presented in unit-dose or multi-dose
containers, for example sealed ampoules and vials, and may be
stored in a freeze-dried (lyophilized) condition requiring only the
addition of the sterile liquid carrier, for example water for
injection, immediately prior to use. Extemporaneous injection
solutions and suspensions are prepared from sterile powders,
granules and tablets of the kind previously described. Preferred
unit dosage formulations are those containing a daily dose or unit
daily sub-dose, as herein above recited, or an appropriate fraction
thereof, of the active ingredient.
[0188] It should be understood that in addition to the ingredients
particularly mentioned above the formulations of this invention may
include other agents conventional in the art having regard to the
type of formulation in question, for example those suitable for
oral administration may include flavoring agents.
[0189] Compounds of the invention can also be formulated to provide
controlled release of the active ingredient to allow less frequent
dosing or to improve the pharmacokinetic or toxicity profile of the
active ingredient. Accordingly, the invention also provided
compositions comprising one or more compounds of the invention
formulated for sustained or controlled release.
[0190] The effective dose of an active ingredient depends at least
on the nature of the condition being treated, toxicity, whether the
compound is being used prophylactically (lower doses) or against an
active viral infection, the method of delivery, and the
pharmaceutical formulation, and will be determined by the clinician
using conventional dose escalation studies. The effective dose can
be expected to be from about 0.0001 to about 100 mg/kg body weight
per day; typically, from about 0.01 to about 10 mg/kg body weight
per day; more typically, from about 0.01 to about 5 mg/kg body
weight per day; most typically, from about 0.05 to about 0.5 mg/kg
body weight per day. For example, the daily candidate dose for an
adult human of approximately 70 kg body weight will range from 1 mg
to 1000 mg, preferably between 5 mg and 500 mg, and may take the
form of single or multiple doses.
[0191] In yet another embodiment, the present application discloses
pharmaceutical compositions comprising a compound of Formula I or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier or exipient.
Routes of Administration
[0192] One or more compounds of the invention (herein referred to
as the active ingredients) are administered by any route
appropriate to the condition to be treated.
[0193] Suitable routes include oral, rectal, nasal, topical
(including buccal and sublingual), vaginal and parenteral
(including subcutaneous, intramuscular, intravenous, intradermal,
intrathecal and epidural), and the like. It will be appreciated
that the preferred route may vary with for example the condition of
the recipient. An advantage of the compounds of this invention is
that they are orally bioavailable and can be dosed orally.
Combination Therapy, Including HCV Combination Therapy
[0194] In another embodiment, the compounds of the present
invention may be combined with one or more active agent.
Non-limiting examples of suitable combinations include combinations
of one or more compounds of the present invention with one or more
interferons, ribavirin or its analogs, HCV NS3 protease inhibitors,
alpha-glucosidase 1 inhibitors, hepatoprotectants, nucleoside or
nucleotide inhibitors of HCV NS5B polymerase, non-nucleoside
inhibitors of HCV NS5B polymerase, HCV NS5A inhibitors, TLR-7
agonists, cyclophillin inhibitors, HCV IRES inhibitors,
pharmacokinetic enhancers, and other drugs for treating HCV.
[0195] More specifically, one or more compounds of the present
invention may be combined with one or more compounds selected from
the group consisting of
[0196] 1) interferons, e.g., pegylated rIFN-alpha 2b (PEG-Intron),
pegylated rIFN-alpha 2a (Pegasys), rIFN-alpha 2b (Intron A),
rIFN-alpha 2a (Roferon-A), interferon alpha (MOR-22, OPC-18,
Alfaferone, Alfanative, Multiferon, subalin), interferon alfacon-1
(Infergen), interferon alpha-n1 (Wellferon), interferon alpha-n3
(Alferon), interferon-beta (Avonex, DL-8234), interferon-omega
(omega DUROS, Biomed 510), albinterferon alpha-2b (Albuferon), IFN
alpha XL, BLX-883 (Locteron), DA-3021, glycosylated interferon
alpha-2b (AVI-005), PEG-Infergen, PEGylated interferon lambda
(PEGylated IL-29), and belerofon,
[0197] 2) ribavirin and its analogs, e.g., ribavirin (Rebetol,
Copegus), and taribavirin (Viramidine),
[0198] 3) HCV NS3 protease inhibitors, e.g., boceprevir
(SCH-503034, SCH-7), telaprevir (VX-950), VX-813, TMC-435
(TMC435350), ABT-450, BI-201335, BI-1230, MK-7009, SCH-900518,
VBY-376, VX-500, GS-9256, GS-9451, BMS-790052, BMS-605339,
PHX-1766, AS-101, YH-5258, YH5530, YH5531, and ITMN-191
(R-7227),
[0199] 4) alpha-glucosidase 1 inhibitors, e.g., celgosivir
(MX-3253), Miglitol, and UT-231B,
[0200] 5) hepatoprotectants, e.g., emericasan (IDN-6556), ME-3738,
GS-9450 (LB-84451), silibilin, and MitoQ,
[0201] 6) nucleoside or nucleotide inhibitors of HCV NS5B
polymerase, e.g., R1626, R7128 (R4048), IDX184, IDX-102, PSI-7851,
BCX-4678, valopicitabine (NM-283), and MK-0608,
[0202] 7) non-nucleoside inhibitors of HCV NS5B polymerase,
filibuvir (PF-868554), ABT-333, ABT-072, BI-207127, VCH-759,
VCH-916, JTK-652, MK-3281, VBY-708, VCH-222, A848837, ANA-598,
GL60667, GL59728, A-63890, A-48773, A-48547, BC-2329, VCH-796
(nesbuvir), GSK625433, BILN-1941, XTL-2125, and GS-9190,
[0203] 8) HCV NS5A inhibitors, e.g., AZD-2836 (A-831), AZD-7295
(A-689), and BMS-790052,
[0204] 9) TLR-7 agonists, e.g., imiquimod, 852A, GS-9524, ANA-773,
ANA-975, AZD-8848 (DSP-3025), PF-04878691, and SM-360320,
[0205] 10) cyclophillin inhibitors, e.g., DEBIO-025, SCY-635, and
NIM811,
[0206] 11) HCV IRES inhibitors, e.g., MC1-067,
[0207] 12) pharmacokinetic enhancers, e.g., BAS-100, SPI-452,
PF-4194477, TMC-41629, GS-9350, GS-9585, and roxythromycin,
[0208] 13) other drugs for treating HCV, e.g., thymosin alpha 1
(Zadaxin), nitazoxanide (Alinea, NTZ), BIVN-401 (virostat), PYN-17
(altirex), KPE02003002, actilon (CPG-10101), GS-9525, KRN-7000,
civacir, G1-5005, XTL-6865, BIT225, PTX-111, ITX2865, TT-033i, ANA
971, NOV-205, tarvacin, EHC-18, VGX-410C, EMZ-702, AVI 4065,
BMS-650032, BMS-791325, Bavituximab, MDX-1106 (ONO-4538),
Oglufanide, FK-788, and VX-497 (merimepodib);
[0209] 14) mevalonate decarboxylase antagonists, e.g., statins,
HMGCoA synthase inhibitors (e.g., hymeglusin), squalene synthesis
inhibitors (e.g., zaragozic acid);
[0210] 15) angiotensin II receptor antagonists, e.g., losartan,
irbesartan, olmesartan, candesartan, valsartan, telmisartan,
eprosartan;
[0211] 16) angiotensin-converting enzyme inhibitors, e.g.,
captopril, zofenopril, enalapril, ramipril, quinapril, perindopril,
lisinopril, benazepril, fosinopril;
[0212] 17) other anti-fibrotic agents, e.g., amiloride and
[0213] 18) endothelin antagonists, e.g. bosentan and
ambrisentan.
[0214] In yet another embodiment, the present application discloses
pharmaceutical compositions comprising a compound of the present
invention, or a pharmaceutically acceptable salt thereof, in
combination with at least one additional active agent, and a
pharmaceutically acceptable carrier or excipient. In yet another
embodiment, the present application provides a combination
pharmaceutical agent with two or more therapeutic agents in a
unitary dosage form. Thus, it is also possible to combine any
compound of the invention with one or more other active agents in a
unitary dosage form.
[0215] The combination therapy may be administered as a
simultaneous or sequential regimen. When administered sequentially,
the combination may be administered in two or more
administrations.
[0216] Co-administration of a compound of the invention with one or
more other active agents generally refers to simultaneous or
sequential administration of a compound of the invention and one or
more other active agents, such that therapeutically effective
amounts of the compound of the invention and one or more other
active agents are both present in the body of the patient.
[0217] Co-administration includes administration of unit dosages of
the compounds of the invention before or after administration of
unit dosages of one or more other active agents, for example,
administration of the compounds of the invention within seconds,
minutes, or hours of the administration of one or more other active
agents. For example, a unit dose of a compound of the invention can
be administered first, followed within seconds or minutes by
administration of a unit dose of one or more other active agents.
Alternatively, a unit dose of one or more other active agents can
be administered first, followed by administration of a unit dose of
a compound of the invention within seconds or minutes. In some
cases, it may be desirable to administer a unit dose of a compound
of the invention first, followed, after a period of hours (e.g.,
1-12 hours), by administration of a unit dose of one or more other
active agents. In other cases, it may be desirable to administer a
unit dose of one or more other active agents first, followed, after
a period of hours (e.g., 1-12 hours), by administration of a unit
dose of a compound of the invention.
[0218] The combination therapy may provide "synergy" and
"synergistic effect", i.e. the effect achieved when the active
ingredients used together is greater than the sum of the effects
that results from using the compounds separately. A synergistic
effect may be attained when the active ingredients are: (1)
co-formulated and administered or delivered simultaneously in a
combined formulation; (2) delivered by alternation or in parallel
as separate formulations; or (3) by some other regimen. When
delivered in alternation therapy, a synergistic effect may be
attained when the compounds are administered or delivered
sequentially, e.g., in separate tablets, pills or capsules, or by
different injections in separate syringes. In general, during
alternation therapy, an effective dosage of each active ingredient
is administered sequentially, i.e. serially, whereas in combination
therapy, effective dosages of two or more active ingredients are
administered together.
[0219] As will be appreciated by those skilled in the art, when
treating a viral infection such as HCV, such treatment may be
characterized in a variety of ways and measured by a variety of
endpoints. The scope of the present invention is intended to
encompass all such characterizations.
SYNTHETIC EXAMPLES
[0220] Certain abbreviations and acronyms are used in describing
the experimental details. Although most of these would be
understood by one skilled in the art, Table 1 contains a list of
many of these abbreviations and acronyms.
TABLE-US-00001 TABLE 1 List of abbreviations and acronyms.
Abbreviation Meaning Ac acetyl ACN acetonitrile AIBN
2,2'-azobis(2-methylpropionitrile) BINAP
2,2'-bis(diphenylphosphino)-1,1'-binaphthyl Bn benzyl BnBr
benzylbromide BSA bis(trimethylsilyl)acetamide BzCl benzoyl
chloride CDI carbonyl diimidazole DABCO
1,4-diazabicyclo[2.2.2]octane dba dibenzylideneacetone DBN
1,5-diazabicyclo[4.3.0]non-5-ene DDQ
2,3-dichloro-5,6-dicyano-1,4-benzoquinone DBU
1,5-diazabicyclo[5.4.0]undec-5-ene DCA dichloroacetamide DCC
dicyclohexylcarbodiimide DCE 1,2-dichloroethane DCM dichloromethane
deg degrees DIAD di-isopropylazodicarboxylate DIEA
N,N-di-isopropylethylamine DMAP 4-dimethylaminopyridine DME
1,2-dimethoxyethane DMTCl dimethoxytrityl chloride DMSO
dimethylsulfoxide DMTr 4,4'-dimethoxytrityl DMF dimethylformamide
EtOAc ethyl acetate ES, ESI electrospray ionization HMDS
hexamethyldisilazane HPLC high pressure liquid chromatography LC
liquid chromatography LDA lithium diisopropylamide LRMS low
resolution mass spectrum MCPBA meta-chloroperbenzoic acid MeCN
acetonitrile MeOH methanol MMTC mono methoxytrityl chloride m/z or
m/e mass to charge ratio MH.sup.+ mass plus 1 MH.sup.- mass minus 1
MsOH methanesulfonic acid MS or ms mass spectrum NBS
N-bromosuccinimide NMP N-methylpyrrolidine Ph phenyl rt or r.t.
room temperature TBAF tetrabutylammonium fluoride TES triethylsilyl
THF tetrahydrofuran THP tetrahydropyran TMSCl chlorotrimethylsilane
TMSBr bromotrimethylsilane TMSI iodotrimethylsilane TMSOTf
(trimethylsilyl)trifluoromethylsulfonate TEA triethylamine TBA
tributylamine TBAP tributylammonium pyrophosphate TBSCl
t-butyldimethylsilyl chloride TEAB triethylammonium bicarbonate TFA
trifluoroacetic acid TLC or tlc thin layer chromatography Tr
triphenylmethyl Tol 4-methylbenzoyl Turbo 1:1 mixture of
isopropylmagnesium chloride and lithium Grignard chloride xantphos
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene .delta. parts per
million down field from tetramethylsilane
General Schemes
[0221] The compounds of this invention may be synthesized by
several routes with key bond-forming steps as indicated in Schemes
A-C, in which the carboxylate substituent R indicates either a
protecting group such as an alkyl ester (where necessary), or the
free acid itself. Alkyl ester protecting groups are conveniently
removed by saponification with an alkali metal hydroxide in a
protic solvent such as water or an alcohol, and may be facilitated
by use of ethereal solvent mixtures and/or heating. Alternatively
they may be removed by dealkylation through heating with an alkali
metal halide in an aprotic solvent. As will be appreciated,
substituents on Het may be modified subsequent to other
bond-forming steps by, for example, N-oxidation with a typical
oxidant such as metachloroperbenzoic acid in a solvent such as
dichloromethane, O-dealkylation through treatment with a reagent
such as boron tribromide in a solvent such as dichloromethane, or
hydrolysis.
##STR00065##
[0222] The bond between L and Het may be formed by displacement of
X on Het, where X is a leaving group such as a halide, sulfinate,
sulfonate or phosphate moiety. The reaction is conveniently
performed by deprotonation of L-H with a base such as sodium
hydride or potassium hexamethyldisilazide, or is facilitated by the
presence of a tertiary amine; it can be carried out in a variety of
solvents such as THF, dioxane, dichloromethane, NMP, DMF or DMSO
and may be accelerated by heating.
##STR00066##
The bond between R.sup.3 and L may be formed by nucleophilic
displacement of a leaving group X on R.sup.3. The leaving group may
vary widely and includes, but is not limited to, halide,
carboxylate, sulfinate, sulfonate or phosphate moieties, and it may
be generated from the corresponding alcohol in situ through
treatment with reagents such as dialkyl azodicarboxylates. The
reaction may also be facilitated by deprotonation of Het-L-H with a
base such as sodium hydride or potassium hexamethyldisilazide, or
is facilitated by the presence of a tertiary amine; it can be
carried out in a variety of solvents such as THF, dioxane,
dichloromethane, NMP, DMF or DMSO and may be accelerated by
heating.
##STR00067##
[0223] The starting material in Scheme A may be synthesized as
depicted in Scheme C. Substituted 3-aminothiophenes II may be
generated by reductive amination of Y--R.sup.3-L-R (where Y
indicates an aldehyde or ketone and R and R.sup.a depict optional
protecting groups), or by direct alkylation (where Y indicates a
leaving group such as a halide, sulfinate, sulfonate or phosphate
moiety) of the 3-aminothiophene I (see patent application
WO2008/58393). In the latter case the alkylation may be facilitated
by deprotonation of the amine with a base such as sodium hydride or
potassium hexamethyldisilazide, and can be carried out in a variety
of solvents such as THF, dioxane, dichloromethane, NMP, DMF or DMSO
and may be accelerated by heating. In cases where R.sup.3 is
aromatic, the reaction may be catalyzed by Pd (J. Org. Chem., 2000,
65, 1158-1174). Alternatively II may be generated by coupling of an
amine with a 3-iodothiophene IV catalyzed by Pd (J. Org. Chem.,
2000, 65, 1158-1174). The amine II is converted to the amide III by
acylation with a carboxylic acid derivative such as an acyl
chloride or anhydride in the presence of a base such as pyridine or
a tertiary amine in an inert solvent such as dichloromethane.
Alternatively IV may be converted to III directly by amidation
catalyzed by Cu (J. Am. Chem. Soc., 2002, 124, 7421-7428).
[0224] The starting material for Scheme B may be generated in an
analogous fashion, with the leaving group X being generated in a
final step by standard methods from the precursor alcohol.
[0225] The synthesis of iodothiophene IV is illustrated below for
the case where R.sup.1=tBu, and other variants may be synthesized
in analogous fashion:
##STR00068##
[0226] To a solution of
5-(3,3-Dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid (6.2 g, 30
mmol; see U.S. Pat. No. 5,861,421) in THF (100 mL) was added a
solution of nBuLi (2.0 M in pentane, 33 mL, 66 mmol) via an
addition funnel at -78.degree. C. After addition, the reaction was
stirred at -78.degree. C. for 1 h. A solution of I.sub.2 (7.7 g, 30
mmol) in THF (100 mL) was added slowly (ca. 15 min) to the flask.
After a further 10 mins, the reaction was quenched with 1 N HCl (50
mL) and warmed to room temperature. The volatiles were removed in
vacuo and the residue was dissolved in ether (500 mL). The organic
solution was washed with 1 M Na.sub.2S.sub.2O.sub.3 (100
mL.times.2), brine (100 mL) and dried over Na.sub.2SO.sub.4. After
concentrated in vacuo, the residue was purified by silica gel
chromatography (EtOAc/hexanes) to give
5-(3,3-Dimethyl-but-1-ynyl)-3-iodo-thiophene-2-carboxylic acid (5.9
g, 65%) as a white solid.
##STR00069##
[0227] To a solution of
5-(3,3-Dimethyl-but-1-ynyl)-3-iodo-thiophene-2-carboxylic acid (1.0
g, 3.0 mmol) and DMF (20 .mu.L) in dry dichloromethane (10 mL) was
added oxalyl chloride (508 .mu.L, 6.0 mmol) at room temperature.
After stirring at room temperature for 90 min, the reaction was
concentrated in vacuo to remove volatiles. The residue was
dissolved in pyridine (5 mL) and methanol (5 mL) and stirred for 2
h. The volatiles were removed in vacuo and the residue was
participated between ether (150 mL) and saturated NH.sub.4Cl
solution (50 mL). The organic layer was washed with saturated
NH.sub.4Cl solution (50 mL) and dried over Na.sub.2SO.sub.4. After
concentration in vacuo, the residue was purified by silica gel
chromatography (EtOAc/hexanes) to give the desired product (835 mg,
80%).
EXPERIMENTALS
Example 31
Compound 31--Synthesis of
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester
##STR00070##
##STR00071##
[0229]
4-[[5-(3,3-dimethyl-but-1-ynyl)-2-methoxycarbonyl-thiophen-3-yl]-(4-
-trans-methyl-cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic
acid tert-butyl ester (1.5 g, 2.73 mmol) was dissolved in ACN (10
mL). To the solution was added a solution of lithium hydroxide (253
mg, 11.01 mmol) in water (10 mL). The reaction was stirred at room
temperature for 6 hours. The reaction was complete as determined by
LC/MS. The pH was adjusted to 5 with 1N HCl in water. The product
was extracted with ethyl acetate (3.times.10 mL). The combined
organics were dried with sodium sulfate, filtered and were
concentrated under reduced pressure and
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid tert-butyl
ester (1.35 g, 93%) was recovered as a white solid.
[0230]
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-m-
ethyl-cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
tert-butyl ester (1.35 g, 2.53 mmol) was dissolved in 4N HCl in
dioxane (6 mL, 24 mmol). The reaction was stirred at room
temperature for 0.5 hours and found to be complete as determined by
LC/MS. The reaction was concentrated under reduced pressure.
[0231] The HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid (100 mg, 0.2 mmol) was
suspended in ACN (1 mL) and a saturated aqueous solution of
NaHCO.sub.3 (1 mL). After 15 minutes,
4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester
chloroformate (1.0 mmol) was added as a solution in THF (1 mL). The
reaction was stirred at room temperature for 1 hour. The reaction
was found to be complete by LC/MS. Boc protected
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester was purified by HPLC to afford a white solid
(76 mg, 53%).
[0232] LC/MS (m/z): 658 [M+1], 558 [M-99]
[0233] Retention time: 2.64 min
[0234] LC: Thermo Finnigan PDA Detector
[0235] MS: Thermo Scientific LCQ Fleet
[0236] Column: Phenomenex Gemini-nx 3u C18 110A 30.times.3 mm
[0237] Solvents: Acetonitrile with 0.1% trifluoroacetic acid, Water
with 0.1% trifluoroacetic acid
[0238] Gradient: 0 min-3.1 min 2%-100% ACN, 3.1 min-3.75 min 100%
ACN
[0239] Boc protected
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester (35 mg, 0.05 mmol) was dissolved in 4N HCl in
dioxane (2 mL, 8.0 mmol). The reaction was stirred at room
temperature for 1 hour. The reaction was complete as determined by
LC/MS. The reaction was concentrated under reduced pressure. The
HCl salt of
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester (31 mg, 94%) was found as a white solid.
[0240] LC/MS (m/z): 558 [M+1]
[0241] Retention time: 2.14 min
[0242] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Synthesis of 4-Hydroxy-piperidine-1-carboxylic acid tert-butyl
ester chloroformate
##STR00072##
[0244] To a solution of 4-hydroxy-piperidine-1-carboxylic acid
tert-butyl ester (201 mg, 1.0 mmol) in THF (2 mL) was added a 20%
solution of phosgene in toluene (358 .mu.L, 1.7 mmol). The reaction
was stirred at room temperature for 1 hour. The reaction was
concentrated under reduced pressure.
Method B
Example 32
Compound 32--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-[4-(t-
etrahydro-pyran-4-yloxycarbonylamino)-cyclohexyl]-amino}-thiophene-2-carbo-
xylic acid
##STR00073##
##STR00074##
[0246]
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-
-(4-oxo-cyclohexyl)-amino]-thiophene-2-carboxylic acid methyl ester
(5.00 g, 10.93 mmol) and ammonium acetate (8.42 g, 109.30 mmol)
were dissolved in MeOH (100 mL) under an atmosphere of nitrogen. To
the reaction was added 4 .ANG. molecular sieves, powdered (500 mg).
After 30 min, sodium triacetoxyborohydride (3.46 g, 16.40 mmol) was
added in 4 portions. The reaction was stirred for 18 h until found
complete by LC/MS. The reaction was filtered through a pad of
celite followed by a MeOH wash and was concentrated.
3-[(4-amino-cyclohexyl)-(4-trans-methyl-cyclohexanecarbonyl)-amino]-5-(3,-
3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid methyl ester
(3.35 g, 67%) was purified by HPLC to afford a white solid.
[0247]
3-[(4-amino-cyclohexyl)-(4-trans-methyl-cyclohexanecarbonyl)-amino]-
-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid methyl
ester (100 mg, 0.22 mmol) was suspended in ACN (1 mL) and a
saturated aqueous solution of NaHCO.sub.3 (1 mL). After 15 minutes,
tetrahydro-pyran-4-ol chloroformate (0.65 mmol) as prepared in a
similar fashion to 4-hydroxy-piperidine-1-carboxylic acid
tert-butyl ester chloroformate except that tetrahydro-pyran-4-ol
was used instead of hydroxy-piperidine-1-carboxylic acid tert-butyl
ester was added as a solution in THF (1 mL). The reaction was
stirred at room temperature for 1.5 hours. The reaction was
complete by LC/MS.
5-(3,3-dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-[4-(t-
etrahydro-pyran-4-yloxycarbonylamino)-cyclohexyl]-amino}-thiophene-2-carbo-
xylic acid methyl ester was extracted with EtOAc (3.times.5 mL).
The combined organics were dried with sodium sulfate, filtered and
were concentrated under reduced pressure.
[0248]
5-(3,3-dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-
-[4-(tetrahydro-pyran-4-yloxycarbonylamino)-cyclohexyl]-amino}-thiophene-2-
-carboxylic acid methyl ester was dissolved in ACN (1 mL). To the
reaction was added a solution of lithium hydroxide (25 mg, 1.1
mmol) in water (1 mL). The reaction was stirred at room temperature
for 2.5 hours. The reaction was complete by LC/MS.
5-(3,3-dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-[4-(t-
etrahydro-pyran-4-yloxycarbonylamino)-cyclohexyl]-amino}-thiophene-2-carbo-
xylic acid (50 mg, 40% over 2 steps) was purified by HPLC to afford
a white solid.
[0249] LC/MS (m/z): 448 [M-124]
[0250] Retention time: 2.46 min
[0251] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 33
Compound 33--Synthesis of
3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-met-
hyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
5-oxo-pyrrolidin-3-(S)-yl ester
##STR00075##
[0253]
3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-tra-
ns-methyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
5-oxo-pyrrolidin-3-(S)-yl ester was prepared in a similar fashion
to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-pyrro-
lidin-3-(S)-yl-amino]-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid and
4-(S)-hydroxy-pyrrolidin-2-one was used instead of
4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester.
[0254] LC/MS (m/z): 544 [M+1]
[0255] Retention time: 1.98 min
[0256] LC: Thermo Finnigan PDA Detector
[0257] MS: Thermo Scientific LCQ Fleet
[0258] Column: Phenomenex Gemini-nx 3u C18 110A 30.times.3 mm
[0259] Solvents: Acetonitrile with 0.1% trifluoroacetic acid, Water
with 0.1% trifluoroacetic acid
[0260] Gradient: 0 min-3.1 min 2%-100% ACN, 3.1 min-3.75 min 100%
ACN
5-(3,3-Dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-pyrrol-
idin-3-(S)-yl-amino]-thiophene-2-carboxylic acid was synthesized as
follows
a) Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-iodo-thiophene-2-carboxylic acid
methyl ester
##STR00076##
[0262] A mixture of 5-Bromo-thiophene-2-carboxylic acid ethyl ester
(7 g, 30 mmol), copper iodide (1.2 g, 6 mmol), triethylamine (20
mL) in DMF (100 mL) was degassed in a 350 mL pressure bottle. Then
tris(dibenzylideneacetone)dipalladium(0) (2.1 g, 3 mmol) and
3,3-dimethyl-but-1-yne (18.3 mL, 150 mmol) were added and heated at
80 degree for 3 hours. The reaction mixture was filtered on celite
and washed with ethyl acetate. The solution was diluted with water
and extracted twice with ethyl acetate. The organic phases were
combined and washed with water. After drying and concentration, the
crude residue was purified by flash chromatography to yield 6.9 g
(95%) of 5-(3,3-Dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid
ethyl ester as a yellow oil.
[0263] A solution of
5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid ethyl ester
(6.9 g) in THF (100 mL) was added LiOH (1.5N, 100 mL). The mixture
was stirred at room temperature for 4 hours. Acidified reaction
with HCl to pH=2, then remove volatiles under vacuo. The resulting
beige color solid was collected by filtration, washed with water
then dried overnight to give 6.2 g of product which was used
without further purification.
[0264] To a solution of
5-(3,3-Dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid (6.2 g, 30
mmol; see U.S. Pat. No. 5,861,421) in THF (100 mL) was added a
solution of nBuLi (2.0 M in pentane, 33 mL, 66 mmol) via an
addition funnel at -78.degree. C. After addition, the reaction was
stirred at -78.degree. C. for 1 h. A solution I.sub.2 (7.7 g, 30
mmol) in THF (100 mL) was added slowly (ca. 15 min) to the flask.
After a further 10 mins, the reaction was quenched with 1 N HCl (50
mL) and warmed to room temperature. The volatiles were removed in
vacuo and the residue was dissolved in ether (500 mL). The organic
solution was washed with 1 M Na.sub.2S.sub.2O.sub.3 (100
mL.times.2), brine (100 mL) and dried over Na.sub.2SO.sub.4. After
concentrated in vacuo, the residue was purified by silica gel
chromatography (EtOAc/hexanes) to give
5-(3,3-Dimethyl-but-1-ynyl)-3-iodo-thiophene-2-carboxylic acid (5.9
g, 65%) as a white solid.
[0265] To a solution of
5-(3,3-dimethyl-but-1-ynyl)-3-iodo-thiophene-2-carboxylic acid (1.0
g, 3.0 mmol) and DMF (20 .mu.L) in dry dichloromethane (10 mL) was
added oxalyl chloride (508 .mu.L, 6.0 mmol) at room temperature.
After stirring at room temperature for 90 min, the reaction was
concentrated in vacuo to remove volatiles. The residue was
dissolved in pyridine (5 mL) and methanol (5 mL) and stirred for 2
h. The volatiles were removed in vacuo and the residue was
participated between ether (150 mL) and saturated NH.sub.4Cl
solution (50 mL). The organic layer was washed with saturated
NH.sub.4Cl solution (50 mL) and dried over Na.sub.2SO.sub.4. After
concentration in vacuo, the residue was purified by silica gel
chromatography (EtOAc/hexanes) to give the desired product (835 mg,
80%).
b) Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-methyl-cyclohexanecarbonyl)-pyrrolidin--
3S-yl-amino]-thiophene-2-carboxylic acid methyl ester
##STR00077##
[0267] A mixture of
5-(3,3-dimethyl-but-1-ynyl)-3-iodo-thiophene-2-carboxylic acid
methyl ester (0.5 g, 1.5 mmol), palladium acetate (0.015 g, 0.32
mmol), BINAP (0.009 g, 0.15 mmol), cesium carbonate (1.2 g, 4.5
mmol) and (3S)-Amino-pyrrolidine-1-carboxylic acid tert-butyl ester
(0.252 g, 1.01 mmol) in toluene (8 mL) was heated to 110.degree. C.
for 8 h. The reaction was diluted with ethyl acetate filtered
through a Celite pad and purified by silica gel chromatography to
give
3-[5-(3,3-dimethyl-but-1-ynyl)-2-methoxycarbonyl-thiophen-3S-ylamino]-pyr-
rolidine-1 carboxylic acid tert-butyl ester in 70% yield.
[0268] To a cooled (0.degree. C.) THF (3 mL) solution of
3-[5-(3,3-dimethyl-but-1-ynyl)-2-methoxycarbonyl-thiophen-3S-ylamino]-pyr-
rolidine-1-carboxylic acid tert-butyl ester (1.0 mmol) was first
added KHMDS (1.0 mmol, 0.5 M in toluene), followed by neat
trans-4-methyl-cyclohexanecarbonyl chloride (0.2 mL, 1.24 mmol).
The reaction was warmed slowly to room temperature and quenched
with saturated ammonium chloride solution. The reaction mixture was
diluted with EtOAc, washed with water, brine, dried over sodium
sulfate, filtrated and concentrated. The crude was then diluted
with EtOAc (10 mL), treated with 4M HCl in dioxane (0.5 mL) and
heated to 50.degree. C. for 30 min. The reaction mixture was cooled
to room temperature concentrated and purified by silica gel column
chromatography to give the title compound in 70% yield.
[0269]
5-(3,3-Dimethyl-but-1-ynyl)-3-[(4-methyl-cyclohexanecarbonyl)-pyrro-
lidin-3S-yl-amino]-thiophene-2-carboxylic acid methyl ester (0.3 g,
0.7 mmol) in a 3:2:1 mixture of THF:MeOH:water (5 mL) was treated
with lithium hydroxide monohydrate (0.69 g, 1.65 mmol) and heated
to 60.degree. C. for 1 hour. The organic volatiles were evaporated
under reduced pressure and the crude material was purified by
reverse-phase HPLC to afford
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-pyrro-
lidin-3-(S)-yl-amino]-thiophene-2-carboxylic acid in 60% yield.
Example 34
Compound 34--Synthesis of
3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-met-
hyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
5-oxo-pyrrolidin-3-(R)-yl ester
##STR00078##
[0271]
3-(S)-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-tra-
ns-methyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
5-oxo-pyrrolidin-3-(R)-yl ester was prepared in a similar fashion
to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-pyrro-
lidin-3-(S)-yl-amino]-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid and
4-(R)-hydroxy-pyrrolidin-2-one was used instead of
4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester.
[0272] LC/MS (m/z): 544 [M+1]
[0273] Retention time: 2.18 min
[0274] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 35
Compound 35--Synthesis of
3-(S)-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-methyl-cy-
clohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
hexahydro-furo[2,3-b]furan-3-(R)-yl ester
##STR00079##
[0276]
3-(S)-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-met-
hyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
hexahydro-furo[2,3-b]furan-3-(R)-yl ester was prepared in a similar
fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-pyrro-
lidin-3-(S)-yl-amino]-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid and 4-nitro-benzoic
acid hexahydro-furo[2,3-b]furan-3-(R)-yl ester was used in place of
the chloroformate of 4-hydroxy-piperidine-1-carboxylic acid
tert-butyl ester.
[0277] LC/MS (m/z): 572.92 [M+1], 595.15 [M+Na.sup.+]
[0278] Retention time: 2.48 min
[0279] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 36
Compound 36--Synthesis of
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-methyl-cycloh-
exanecarbonyl)-amino]-piperidine-1-carboxylic acid
hexahydro-furo[2,3-b]furan-3-(R)-yl ester
##STR00080##
[0281]
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
hexahydro-furo[2,3-b]furan-3-(R)-yl ester was prepared in a similar
fashion to
5-(3,3-dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-[4-(t-
etrahydro-pyran-4-yloxycarbonylamino)-cyclohexyl]-amino}-thiophene-2-carbo-
xylic acid using method B, except the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-methyl-cyclohexanecarbonyl)-piperidin-4-
-yl-amino]-thiophene-2-carboxylic acid methyl ester was used
instead of
3-[(4-amino-cyclohexyl)-(4-trans-methyl-cyclohexanecarbonyl)-amino]-5-(3,-
3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid methyl ester,
and 4-nitro-benzoic acid hexahydro-furo[2,3-b]furan-3-(R)-yl ester
was used in place of the chloroformate of
tetrahydro-pyran-4-ol.
[0282] LC/MS (m/z): 587.01 [M+1], 609.20 [M+Na.sup.+]
[0283] Retention time: 2.40 min
[0284] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 37
Compound 37--Synthesis of
3-(R)-{3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-tr-
ans-methyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carbonyloxy}-piperidi-
ne-1-carboxylic acid tert-butyl ester
##STR00081##
[0286]
3-(R)-{3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-
-(4-trans-methyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carbonyloxy}-pi-
peridine-1-carboxylic acid tert-butyl ester was prepared in a
similar fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-pyrro-
lidin-3-(S)-yl-amino]-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid and
3-(R)-hydroxy-piperidine-1-carboxylic acid tert-butyl ester was
used instead of 4-hydroxy-piperidine-1-carboxylic acid tert-butyl
ester.
[0287] LC/MS (m/z): 545 [M-98]
[0288] Retention time: 2.26 min
[0289] LC: Thermo Finnigan PDA Detector
[0290] MS: Thermo Scientific LCQ Fleet
[0291] Column: Phenomenex Gemini-nx 3u C18 110A 30.times.3 mm
[0292] Solvents: Acetonitrile with 0.1% trifluoroacetic acid, Water
with 0.1% trifluoroacetic acid
[0293] Gradient: 0 min-3.1 min 2%-100% ACN, 3.1 min-3.75 min 100%
ACN
Example 38
Compound 38--Synthesis of
3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-met-
hyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
piperidin-3-(R)-yl ester
##STR00082##
[0295]
3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-tra-
ns-methyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
piperidin-3-(R)-yl ester was prepared in a similar fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-pyrro-
lidin-3-(S)-yl-amino]-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid and
3-(R)-hydroxy-piperidine-1-carboxylic acid tert-butyl ester was
used instead of 4-hydroxy-piperidine-1-carboxylic acid tert-butyl
ester.
[0296] LC/MS (m/z): 544 [M+1]
[0297] Retention time: 2.18 min
[0298] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 39
Compound 39--Synthesis of
4-{3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans--
methyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carbonyloxy}-piperidine-1-
-carboxylic acid tert-butyl ester
##STR00083##
[0300]
4-{3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4--
trans-methyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carbonyloxy}-piperi-
dine-1-carboxylic acid tert-butyl ester was prepared in a similar
fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-meth-
yl-cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-pyrro-
lidin-3-(S)-yl-amino]-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid.
[0301] LC/MS (m/z): 644 [M+1], 544 [M-99]
[0302] Retention time: 2.56 min
[0303] LC: Thermo Finnigan PDA Detector
[0304] MS: Thermo Scientific LCQ Fleet
[0305] Column: Phenomenex Gemini-nx 3u C18 110A 30.times.3 mm
[0306] Solvents: Acetonitrile with 0.1% trifluoroacetic acid, Water
with 0.1% trifluoroacetic acid
[0307] Gradient: 0 min-3.1 min 2%-100% ACN, 3.1 min-3.75 min 100%
ACN
Example 40
Compound 40--Synthesis of
3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-met-
hyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
piperidin-4-yl ester
##STR00084##
[0309]
3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-tra-
ns-methyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
piperidin-4-yl ester was prepared in a similar fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-pyrro-
lidin-3-(S)-yl-amino]-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid.
[0310] LC/MS (m/z): 544 [M+1]
[0311] Retention time: 2.09 min
[0312] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 41
Compound 41--Synthesis of
3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-met-
hyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
tetrahydro-pyran-4-yl ester
##STR00085##
[0314]
3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-tra-
ns-methyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
tetrahydro-pyran-4-yl ester was prepared in a similar fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-pyrro-
lidin-3-(S)-yl-amino]-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid and
tetrahydro-pyran-4-ol was used instead of
4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester.
[0315] LC/MS (m/z): 544 [M]
[0316] Retention time: 2.57 min
[0317] LC: Thermo Finnigan PDA Detector
[0318] MS: Thermo Scientific LCQ Fleet
[0319] Column: Phenomenex Gemini-nx 3u C18 110A 30.times.3 mm
[0320] Solvents: Acetonitrile with 0.1% trifluoroacetic acid, Water
with 0.1% trifluoroacetic acid
[0321] Gradient: 0 min-3.1 min 2%-100% ACN, 3.1 min-3.75 min 100%
CAN
Example 42
Compound 42--Synthesis of
3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-met-
hyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
tetrahydro-furan-3-(R)-yl ester
##STR00086##
[0323]
3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-tra-
ns-methyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
tetrahydro-furan-3-(R)-yl ester was prepared in a similar fashion
to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-pyrro-
lidin-3-(S)-yl-amino]-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid and
tetrahydro-furan-3-(R)-ol was used instead of
4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester.
[0324] LC/MS (m/z): 531 [M+1]
[0325] Retention time: 2.21 min
[0326] LC: Thermo Finnigan PDA Detector
[0327] MS: Thermo Scientific LCQ Fleet
[0328] Column: Phenomenex Gemini-nx 3u C18 110A 30.times.3 mm
[0329] Solvents: Acetonitrile with 0.1% trifluoroacetic acid, Water
with 0.1% trifluoroacetic acid
[0330] Gradient: 0 min-3.1 min 2%-100% ACN, 3.1 min-3.75 min 100%
ACN
Example 43
Compound 43--Synthesis of
3-(S)-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-met-
hyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
2-oxo-pyrrolidin-3-(R)-yl ester
##STR00087##
[0332]
3-(S)-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-tra-
ns-methyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxylic acid
2-oxo-pyrrolidin-3-(R)-yl ester was prepared in a similar fashion
to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-pyrro-
lidin-3-(S)-yl-amino]-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid and
3-(R)-hydroxy-pyrrolidin-2-one was used instead of
4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester.
[0333] LC/MS (m/z): 562 [M+19]
[0334] Retention time: 2.18 min
[0335] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 44
Compound 44--Synthesis of
4-{4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-meth-
yl-cyclohexanecarbonyl)-amino]-cyclohexylcarbamoyloxy}-piperidine-1-carbox-
ylic acid tert-butyl ester
##STR00088##
[0337]
4-{4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-tran-
s-methyl-cyclohexanecarbonyl)-amino]-cyclohexylcarbamoyloxy}-piperidine-1--
carboxylic acid tert-butyl ester was prepared in a similar fashion
to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
3-[(4-amino-cyclohexyl)-(4-trans-methyl-cyclohexanecarbonyl)-amino]-5-(3,-
3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid.
[0338] LC/MS (m/z): 670 [M-1]
[0339] Retention time: 2.54 min
[0340] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 45
Compound 45--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-[4-(p-
iperidin-4-yloxycarbonylamino)-cyclohexyl]-amino}-thiophene-2-carboxylic
acid
##STR00089##
[0342]
5-(3,3-Dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-
-[4-(piperidin-4-yloxycarbonylamino)-cyclohexyl]-amino}-thiophene-2-carbox-
ylic acid was prepared in a similar fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
3-[(4-amino-cyclohexyl)-(4-trans-methyl-cyclohexanecarbonyl)-amino]-5-(3,-
3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid.
[0343] LC/MS (m/z): 572 [M+1]
[0344] Retention time: 2.07 min
[0345] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 46
Compound 46--Synthesis of
3-(R)-{4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans--
methyl-cyclohexanecarbonyl)-amino]-cyclohexylcarbamoyloxy}-piperidine-1-ca-
rboxylic acid tert-butyl ester
##STR00090##
[0347]
3-(R)-{4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4--
trans-methyl-cyclohexanecarbonyl)-amino]-cyclohexylcarbamoyloxy}-piperidin-
e-1-carboxylic acid tert-butyl ester was prepared in a similar
fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
3-[(4-amino-cyclohexyl)-(4-trans-methyl-cyclohexanecarbonyl)-amino]-5-(3,-
3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid and
3-(R)-hydroxy-piperidine-1-carboxylic acid tert-butyl ester was
used instead of 4-hydroxy-piperidine-1-carboxylic acid tert-butyl
ester.
[0348] LC/MS (m/z): 670 [M-1]
[0349] Retention time: 2.58 min
[0350] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 47
Compound 47--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-[4-(p-
iperidin-3-(R)-yloxycarbonylamino)-cyclohexyl]-amino}-thiophene-2-carboxyl-
ic acid
##STR00091##
[0352]
5-(3,3-dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-
-[4-(piperidin-3-(R)-yloxycarbonylamino)-cyclohexyl]-amino}-thiophene-2-ca-
rboxylic acid was prepared in a similar fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
3-[(4-amino-cyclohexyl)-(4-trans-methyl-cyclohexanecarbonyl)-amino]-5-(3,-
3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid and
3-(R)-hydroxy-piperidine-1-carboxylic acid tert-butyl ester was
used instead of 4-hydroxy-piperidine-1-carboxylic acid tert-butyl
ester.
[0353] LC/MS (m/z): 572 [M+1]
[0354] Retention time: 2.11 min
[0355] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 48
Compound 48--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-[4-(t-
etrahydro-furan-3-(R)-yloxycarbonylamino)-cyclohexyl]-amino}-thiophene-2-c-
arboxylic acid
##STR00092##
[0357]
5-(3,3-Dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-
-[4-(tetrahydro-furan-3-(R)-yloxycarbonylamino)-cyclohexyl]-amino}-thiophe-
ne-2-carboxylic acid was prepared in a similar fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
3-[(4-amino-cyclohexyl)-(4-trans-methyl-cyclohexanecarbonyl)-amino]-5-(3,-
3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid and
tetrahydro-furan-3-(R)-ol was used instead of
4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester.
[0358] LC/MS (m/z): 434 [M-124]
[0359] Retention time: 2.42 min
[0360] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 49
Compound 49--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-[4-(5-
-oxo-pyrrolidin-3-(S)-yloxycarbonylamino)-cyclohexyl]-amino}-thiophene-2-c-
arboxylic acid
##STR00093##
[0362]
5-(3,3-Dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-
-[4-(5-oxo-pyrrolidin-3-(S)-yloxycarbonylamino)-cyclohexyl]-amino}-thiophe-
ne-2-carboxylic acid was prepared in a similar fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
3-[(4-amino-cyclohexyl)-(4-trans-methyl-cyclohexanecarbonyl)-amino]-5-(3,-
3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid and
4-(S)-hydroxy-pyrrolidin-2-one was used instead of
4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester.
[0363] LC/MS (m/z): 572 [M+1]
[0364] Retention time: 2.08 min
[0365] LC: Thermo Finnigan PDA Detector
[0366] MS: Thermo Scientific LCQ Fleet
[0367] Column: Phenomenex Gemini-nx 3u C18 110A 30.times.3 mm
[0368] Solvents: Acetonitrile with 0.1% trifluoroacetic acid, Water
with 0.1% trifluoroacetic acid
[0369] Gradient: 0 min-3.1 min 2%-100% ACN, 3.1 min-3.75 min 100%
ACN
Example 50
Compound 50--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-[4-(2-
-oxo-pyrrolidin-3-(R)-yloxycarbonylamino)-cyclohexyl]-amino}-thiophene-2-c-
arboxylic acid
##STR00094##
[0371]
5-(3,3-Dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-
-[4-(2-oxo-pyrrolidin-3-(R)-yloxycarbonylamino)-cyclohexyl]-amino}-thiophe-
ne-2-carboxylic acid was prepared in a similar fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
3-[(4-amino-cyclohexyl)-(4-trans-methyl-cyclohexanecarbonyl)-amino]-5-(3,-
3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid and
3-(R)-hydroxy-pyrrolidin-2-one was used instead of
4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester.
[0372] LC/MS (m/z): 447 [M-124]
[0373] Retention time: 2.52 min
[0374] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 51
Compound 51--Synthesis of Boc protected
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-3-(R)-yl ester
##STR00095##
[0376] Boc protected
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-3-(R)-yl ester was prepared in a similar fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that
3-(R)-hydroxy-piperidine-1-carboxylic acid tert-butyl ester was
used instead of 4-hydroxy-piperidine-1-carboxylic acid tert-butyl
ester.
[0377] LC/MS (m/z): 558 [M-99]
[0378] Retention time: 2.64 min
[0379] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 52
Compound 52--Synthesis of
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-3-(R)-yl ester
##STR00096##
[0381]
4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-m-
ethyl-cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-3-(R)-yl ester was prepared in a similar fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that
3-(R)-hydroxy-piperidine-1-carboxylic acid tert-butyl ester was
used instead of 4-hydroxy-piperidine-1-carboxylic acid tea-butyl
ester.
[0382] LC/MS (m/z): 558 [M+1]
[0383] Retention time: 2.15 min
[0384] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 53
Compound 53--Synthesis of
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
tetrahydro-furan-3-(R)-yl ester
##STR00097##
[0386]
4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-m-
ethyl-cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
tetrahydro-furan-3-(R)-yl ester was prepared in a similar fashion
to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl except that tetrahydro-furan-3-(R)-ol was used
instead of 4-hydroxy-piperidine-1-carboxylic acid tert-butyl
ester.
[0387] LC/MS (m/z): 545 [M+1]
[0388] Retention time: 2.28 min
[0389] LC: Thermo Finnigan PDA Detector
[0390] MS: Thermo Scientific LCQ Fleet
[0391] Column: Phenomenex Gemini-nx 3u C18 110A 30.times.3 mm
[0392] Solvents: Acetonitrile with 0.1% trifluoroacetic acid, Water
with 0.1% trifluoroacetic acid
[0393] Gradient: 0 min-3.1 min 2%-100% ACN, 3.1 min-3.75 min 100%
ACN
Example 54
Compound 54--Synthesis of
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
tetrahydro-pyran-4-yl ester
##STR00098##
[0395]
4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-m-
ethyl-cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
tetrahydro-pyran-4-yl ester was prepared in a similar fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl except that tetrahydro-pyran-4-ol was used instead
of 4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester.
[0396] LC/MS (m/z): 559 [M+1]
[0397] Retention time: 2.35 min
[0398] Gradient: 0 min-3.1 min 2%-100% ACN, 3.1 min-3.75 min 100%
ACN
Example 55
Compound 55--Synthesis of
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
5-oxo-pyrrolidin-3-(R)-yl ester
##STR00099##
[0400]
4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-m-
ethyl-cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
5-oxo-pyrrolidin-3-(R)-yl ester was prepared in a similar fashion
to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl except that 4-(R)-hydroxy-pyrrolidin-2-one was used
instead of 4-hydroxy-piperidine-1-carboxylic acid tert-butyl
ester.
[0401] LC/MS (m/z): 433 [M-124]
[0402] Retention time: 2.54 min
[0403] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 56
Compound 56--Synthesis of
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
5-oxo-pyrrolidin-3-(S)-yl ester
##STR00100##
[0405]
4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-m-
ethyl-cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
5-oxo-pyrrolidin-3-(R)-yl ester was prepared in a similar fashion
to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl except that 4-(S)-hydroxy-pyrrolidin-2-one was used
instead of 4-hydroxy-piperidine-1-carboxylic acid tert-butyl
ester.
[0406] LC/MS (m/z): 433 [M-124]
[0407] Retention time: 2.54 min
[0408] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN
Example 57
Compound 57--Synthesis of
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-methyl-cycloh-
exanecarbonyl)-amino]-piperidine-1-carboxylic acid
2-oxo-pyrrolidin-3-(S)-yl ester
##STR00101##
[0410]
4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
2-oxo-pyrrolidin-3-(S)-yl ester was prepared in a similar fashion
to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl except that 3-(S)-Hydroxy-pyrrolidin-2-one was used
instead of 4-hydroxy-piperidine-1-carboxylic acid tert-butyl
ester.
[0411] LC/MS (m/z): 576 [M+19]
[0412] Retention time: 2.16 min
[0413] LC: Thermo Electron Surveyor HPLC
[0414] MS: Finnigan LCQ Advantage MAX Mass Spectrometer
[0415] Column: Phenomenex Polar RP 30 mm.times.4.6 mm
[0416] Solvents: Acetonitrile with 0.1% formic acid, Water with
0.1% formic acid
[0417] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN
Method D
Example 64
Compound 64--Synthesis of
5-(3,3-Dimethyl-but-1-ynyl)-3-[[3-(hexahydro-furo[2,3-b]furan-3-yloxycarb-
onylamino)-1-(S)-methyl-propyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino-
]-thiophene-2-carboxylic acid
##STR00102##
[0419] A mixture of
5-(3,3-dimethyl-but-1-ynyl)-3-[(3-hydroxy-1-(S)-methyl-propyl)-(trans-4-m-
ethyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylic acid TFA
salt (1.40 g, 3.23 mmol) in THF (30 mL) was cooled to 0.degree. C.
and triphenylphosphine (2.53 g, 9.69 mmol) was added followed by
phthalimide (0.713 g, 4.84 mmol). The reaction was stirred until
homogenious then DIAD (1.06 mL, 5.49 mmol) was added. After warming
to rt., the reaction was determined to be complete by LC/MS in 1 h.
The reaction was quenched with CH.sub.3OH (1.0 mL) and the solvent
removed under reduced pressure.
5-(3,3-Dimethyl-but-1-ynyl)-3-[[3-(1,3-dioxo-1,3-dihydro-isoindole-2-yl)--
1-(S)-methyl
propyl]-(trans-4-methyl-cyclohexylcarbonyl)-amino]-thiophene-2-carboxylic
acid methyl ester (1.67 g, 92%) was isolated by silica gel
chromatography as an off-white solid.
[0420] A mixture of
5-(3,3-dimethyl-but-1-ynyl)-3-[[3-(1,3-dioxo-1,3-dihydro-isoindole-2-yl)--
1-(S)-methyl
propyl]-(trans-4-methyl-cyclohexylcarbonyl)-amino]-thiophene-2-carboxylic
acid methyl ester (168 mg, 0.30 mmol) in CH.sub.3OH (2 mL) and
hydrazine (15 .mu.L, 0.45 mmol) was placed in a preheated
80.degree. C. oil bath and stirred for 2 h. The reaction was
determined to be complete by LC/MS. Solvent was removed under
reduced pressure and the crude material was coevaporated with
toluene.
3-[[3-Amino-1-(S)-methyl-propyl)-(trans-4-methyl-cyclohexanecarbonyl)-ami-
no]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid methyl
ester was used as is for the next step without purification.
[0421] A mixture of crude
3-[[3-Amino-1-(S)-methyl-propyl)-(trans-4-methyl-cyclohexanecarbonyl)-ami-
no]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid methyl
ester in ACN (3.0 mL) was treated with carbonic acid
hexahydro-furo[2,3-b]furan-3-yl ester 4-nitro-phenyl ester (0.11 g,
0.375 mmol) followed by diisopropylethyl amine (0.1 mL, 0.625 mmol)
and DMAP (cat). The reaction was stirred for 1 h and quenched with
CH.sub.3OH (1 mL). Solvent was removed under reduced pressure and
the crude material was partitioned between EtOAc, and 1/2 sat
NaHCO.sub.3(aq). The organics were dried over Na.sub.2SO.sub.4,
solids filtered and solvent removed under reduced pressure.
5-(3,3-Dimethyl-but-1-ynyl)-3-[[3-(hexahydro-furo[2,3-b]furan-3-yloxycarb-
onylamino)-1-(S)-methyl-propyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino-
]-thiophene-2-carboxylic acid methyl ester was isolated by reverse
phase HPLC and carried forward wet to the next reaction.
[0422] A mixture of
5-(3,3-Dimethyl-but-1-ynyl)-3-[[3-(hexahydro-furo[2,3-b]furan-3-yloxycarb-
onylamino)-1-(S)-methyl-propyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino-
]-thiophene-2-carboxylic acid methyl ester in THF (2 mL) and
CH.sub.3OH (1 mL) was treated with dissolve LiOH.H.sub.2O (0.1 g,
2.38 mmol) in H.sub.2O (2 mL). The reaction was determined to be
complete after 5 h. The pH was adjusted to 2 with 2N HCl.sub.(aq).
After diluting with CH.sub.3OH (3 mL),
5-(3,3-Dimethyl-but-1-ynyl)-3-[[3-(hexahydro-furo[2,3-b]furan-3-yloxycarb-
onylamino)-1-(S)-methyl-propyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino-
]-thiophene-2-carboxylic acid TFA salt (0.41 g, 24% 3-steps) was
isolated by reverse phase HPLC as a white solid.
[0423] LC/MS=597.20 (M.sup.++Na)
[0424] Retention time: 3.76 min
[0425] Gradient: 0 min-0.2 min 5% ACN, 0.2 min-3.95 min 5%-100%
ACN, 3.95 min-5.20 min 100% ACN, 5.20 min-5.5 min 100%-5% ACN, 5.5
min-6 min 5% ACN.
Example 65
Compound 65--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-[[trans-4-methyl-cyclohexanecarbonyl)-[1-(S-
)-methyl-3-(tetrahydrofuran-3-yloxycarbonylamino)-propyl]-amino]-thiophene-
-2-carboxylic acid
##STR00103##
[0427] A solution of
3-[[3-Amino-1-(S)-methyl-propyl)-(trans-4-methyl-cyclohexanecarbonyl)-ami-
no]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid methyl
ester (0.08 g, 0.146 mmol) in CH.sub.3CN (1.5 mL) was prepared.
3-(R)-Hydroxy-tetrahydrofuranyl-chloroformate (0.132 g, 0.88 mmol),
which was prepared in a manner similar to method G, was added
followed by diisopropylethyl amine (0.25 mL, 1.50 mmol) and DMAP
(cat). After stirring at rt for 30 min., the reaction was
determined to be complete by LC/MS. The reaction was quenched with
H.sub.2O (1 mL) and the organics removed under reduced pressure and
5-(3,3-dimethyl-but-1-ynyl)-3-[[trans-4-methyl-cyclohexanecarbonyl)-[1-(S-
)-methyl-3-(tetrahydrofuran-3-yloxycarbonylamino)-propyl]-amino]-thiophene-
-2-carboxylic acid methyl ester was carried forward without
purification.
[0428] A mixture of
5-(3,3-dimethyl-but-1-ynyl)-3-[[trans-4-methyl-cyclohexanecarbonyl)-[1-(S-
)-methyl-3-(tetrahydrofuran-3-yloxycarbonylamino)-propyl]-amino]-thiophene-
-2-carboxylic acid methyl ester in THF (2 mL) and CH.sub.3OH (1 mL)
was treated with dissolve LiOH.H.sub.2O (0.1 g, 2.38 mmol) in
H.sub.2O (2 mL). The reaction was determined to be complete after 1
h. The pH was adjusted to 2 with 2NHCl.sub.(aq), the reaction
diluted with CH.sub.3OH (3 mL) and
5-(3,3-dimethyl-but-1-ynyl)-3-[[trans-4-methyl-cyclohexanecarbonyl)-[1-(S-
)-methyl-3-(tetrahydrofuran-3-yloxycarbonylamino)-propyl]-amino]-thiophene-
-2-carboxylic acid TFA salt (0.046 g, 47%) was isolated by reverse
phase HPLC as a white solid.
[0429] LC/MS=555.24 (M+Na.sup.+)
[0430] Retention time: 3.65 min
[0431] Gradient: 0 min-0.2 min 5% ACN, 0.2 min-3.95 min 5%-100%
ACN, 3.95 min-5.20 min 100% ACN, 5.20 min-5.5 min 100%-5% ACN, 5.5
min-6 min 5% ACN.
Method E
Example 66
Compound
66--5-(3,3-Dimethyl-but-ynyl)-3-[[4-(hexahydro-furo[2,3-b]furan-3-
-yloxycarbonylamino)-cyclohexyl]-(trans-4-methyl-cyclohexanecarbonyl)-amin-
o]-thiophene-2-carboxylic acid
##STR00104##
##STR00105##
[0433] A solution of
3-[(4-amino-cyclohexyl)-(trans-4-methylcyclohexanecarbonyl)-amino]-5-(3,3-
-dimethyl-but-1ynyl)thiophene-2-carboxylic acid methyl ester (0.40
g, 0.873 mmol) in ACN (5.0 mL) was cooled to 0.degree. C. and
carbonic acid hexahydro-furo[2,3-b]furan-3-yl ester 4-nitro-phenyl
ester (0.295 g, 1.0 mmol) was added, followed by DIEA (391 .mu.L,
2.18 mmol) and DMAP (cat). The reaction mixture was warmed to rt,
and stirred for 1 h. The solvent was removed under reduced pressure
and the crude reaction mixture was partitioned between EtOAc and 2N
K.sub.2CO.sub.3(aq).
[0434] The layers were separated and the organics washed repeatedly
with 2N K.sub.2CO.sub.3(aq). Both epimers of
5-(3,3-Dimethyl-but-ynyl)-3-[[4-(hexahydro-furo[2,3-b]furan-3-yloxycarbon-
ylamino)-cyclohexyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-
-2-carboxylic acid methyl ester were isolated by reverse phase HPLC
and carried forward wet into the next reaction.
[0435] To each of the wet fractions of
5-(3,3-Dimethyl-but-ynyl)-3-[[4-(hexahydro-furo[2,3-b]furan-3-yloxycarbon-
ylamino)-cyclohexyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-
-2-carboxylic acid methyl ester were added THF (0.5 mL), CH.sub.3OH
(0.2 mL), and LiOH (0.021 g, 0.5 mmol). The reactions were allowed
to stir at rt for 2 h. The reactions were determined to be complete
by LC/MS. The reactions were quenched with 2N HCl.sub.(aq) to
pH=ca. 2. Both epimers of
5-(3,3-Dimethyl-but-ynyl)-3-[[4-(hexahydro-furo[2,3-b]furan-3-yloxycarbon-
ylamino)-cyclohexyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-
-2-carboxylic acid TFA salt (fast eluting: 89 mg, 34%, slow eluting
34 mg, 14%) were isolated by reverse phase HPLC as an off-white
solid.
[0436] LC/MS=623.20 (M.sup.++Na)
[0437] Retention time: 3.76 min
[0438] Gradient: 0 min-0.2 min 5% ACN, 0.2 min-3.95 min 5%-100%
ACN, 3.95 min-5.20 min 100% ACN, 5.20 min-5.5 min 100%-5% ACN, 5.5
min-6 min 5% ACN.
Method F
Example 67
Compound
67--3-(S)-{3-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophene-3--
yl](trans-4-methyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1
carbonyloxy}-3-methyl-pyrrolidine-1-carboxylic acid tert-butyl
ester
##STR00106##
[0439] And
Example 68
Compound
68--3-(S)-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]--
(trans-4-methylcyclohexanecarbonyl)-amino]-pyrrolidine-1-carbocylic-acid-3-
-methyl-pyrrolidin-3-yl-ester
##STR00107##
##STR00108##
[0441]
3-(S)-[[5-(3,3-Dimethyl-but-1-ynyl)-2-methoxycarbonyl-thiophene-3-(-
S)-yl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-pyrrolidine-1-carboxyli-
c acid tert-butyl ester (3.10 g, 5.85 mmol) was taken up in
CH.sub.2Cl.sub.2 (50 mL) and 4N HCl in dioxane (25 mL, 100 mmol)
was added in one portion. The reaction was stirred for 2 h and was
determined to be complete by LC/MS. Solvents were removed under
reduced pressure and the crude reaction mixture was co-evaporated
with 3.times.25 mL toluene.
5-(3,3-dimethyl-but-1-ynyl)-3-[[trans-4-methyl-cyclohexanecarbonyl)-pyrro-
lidin-3-(S)-yl-amino]-thiophene-carboxylic acid methyl ester was
carried forward crude into the next step.
[0442] A mixture of
5-(3,3-dimethyl-but-1-ynyl)-3-[[trans-4-methyl-cyclohexanecarbonyl)-pyrro-
lidin-3-(S)-yl-amino]-thiophene-carboxylic acid methyl ester in THF
(25 mL) and CH.sub.3OH (10 mL) was treated with LiOH.H.sub.2O (1.43
g, 34.2 mmol) in H.sub.2O (10 mL). The reaction was determined to
be complete by LC/MS after 2 h. The pH was adjusted to 2 with 2N
HCl.sub.(aq) and the solvents were removed under reduced pressure.
Co-evaporation with CH.sub.3OH, then EtOAc, and finally toluene
removed all H.sub.2O.
5-(3,3-Dimethyl-but-1-ynyl)-3-[[trans-4-methyl-cyclohexanecarbonyl)-pyrro-
lidin-3-(S)-yl-amino]-thiophene-carboxylic acid was carried forward
crude with LiCl salt into the next step.
[0443]
5-(3,3-Dimethyl-but-1-ynyl)-3-[[trans-4-methyl-cyclohexanecarbonyl)-
-pyrrolidin-3-(S)-yl-amino]-thiophene-carboxylic acid (150 mg) was
taken up in ACN (2.0 mL). The heterogeneous solution was allowed to
stir for 5 min, then DIEA (250 .mu.L, 2.3 mmol) and DMAP (cat) was
added sequentially to the solution. The reaction was allowed to
stir for 5 min then
3-chlorocarbonyloxy-3-methyl-pyrrolidine-1-carboxylic acid
tert-butyl ester (.apprxeq.200 mg) prepared in a manner similar to
method G was added in one portion. The reaction was stirred at rt
for 15 min and determined to be complete by LC/MS. Solvent was
removed under reduced pressure. The reaction mixture was
partitioned between EtOAc and 2N HCl.sub.(aq). The organics were
dried over Na.sub.2SO.sub.4, solids filtered and solvent removed
under reduced pressure.
3-{3-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophene-3-yl]-(trans-4-met-
hyl-cyclohexanecarbonyl)-3-(S)-amino]-pyrrolidine-1-carbonyloxy}-3-methyl--
pyrrolidine-1-carboxylic acid tert-butyl ester TFA salt (35 mg,
20%) and
3-{3-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophene-3-yl]-(trans-4-met-
hyl-cyclohexanecarbonyl)-3-(S)-amino]-pyrrolidine-1-carbonyloxy}-3-methyl--
pyrrolidine-1-carboxylic acid TFA salt (11 mg, 7%) were isolated by
reverse phase HPLC as white solids.
3-{3-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophene-3-yl]-(trans-4-meth-
yl-cyclohexanecarbonyl)-3-(S)-amino]-pyrrolidine-1-carbonyloxy}-3-methyl-p-
yrrolidine-1-carboxylic acid tert-butyl ester TFA salt
[0444] LC/MS=544.14 (M.sup.+-Boc)
[0445] Retention time: 4.02 min
[0446] Gradient: 0 min-0.2 min 5% ACN, 0.2 min-3.95 min 5%-100%
ACN, 3.95 min-5.20 min 100% ACN, 5.20 min-5.5 min 100%-5% ACN, 5.5
min-6 min 5% ACN.
3-{3-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophene-3-yl]-(trans-4-meth-
yl-cyclohexanecarbonyl)-3-(S)-amino]-pyrrolidine-1-carbonyloxy}-3-methyl-p-
yrrolidine-1-carboxylic acid TFA salt
[0447] LC/MS=544.10 (M.sup.++1)
[0448] Retention time: 3.36 min
[0449] Gradient: 0 min-0.2 min 5% ACN, 0.2 min-3.95 min 5%-100%
ACN, 3.95 min-5.20 min 100% ACN, 5.20 min-5.5 min 100%-5% ACN, 5.5
min-6 min 5% ACN.
Method G
3-Chlorocarbonyloxy-3-methyl-pyrrolidine-1-carboxylic acid
tert-butyl ester
##STR00109##
[0451] A solution of 3-hydroxy-3-methylpyrrolidine-carboxylic acid
tert-butyl ester (200 mg, 1.00 mmol) in THF (4 mL) was slowly
treated with phosgene 20% in toluene (0.954 mL). After stirring at
rt for 16 h, solvents were removed under reduced pressure and
co-evaporate with CH.sub.2Cl.sub.2. The crude
3-chlorocarbonyloxy-3-methylpyrrolidine-carboxylic acid tert-butyl
ester was used directly in the next step.
Example 69
Compound 69--Synthesis of
3-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(trans-4-methyl--
cyclohexanecarbonyl)-3-(S)-amino]-pyrrolidine-1-carboxylic acid
1-tert-butoxycarbonyl-3-methyl-azetidin-3-yl ester
##STR00110##
[0453]
5-(3,3-Dimethyl-but-1-ynyl)-3-[[trans-4-methyl-cyclohexanecarbonyl)-
-pyrrolidin-3-(S)-yl-amino]-thiophene-carboxylic acid (150 mg) was
taken up in ACN (2.0 mL). The heterogeneous solution was allowed to
stir for 5 min, and then DIEA (250 .mu.L, 2.3 mmol) and DMAP (cat)
were added sequentially. The reaction was allowed to stir for 5 min
then 3-chlorocarbonyloxy-3-methyl-azetidine-1-carboxylic acid
tert-butyl ester (.apprxeq.200 mg) which was prepared in a similar
manner to Method G was added in one portion. The reaction was
stirred at rt for 15 min and determined to be complete by LC/MS.
Solvent was removed under reduced pressure. The reaction mixture
was partitioned between EtOAc and 2N HCl.sub.(aq). The organics
were dried over Na.sub.2SO.sub.4, solids filtered and solvent
removed under reduced pressure.
3-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophene-3-yl]-(trans-4-methyl-
-cyclohexanecarbonyl)-3-(S)-amino]-pyrrolidine-1-carboxylic acid
1-tert-butoxycarbonyl-3-methyl-azetidin-3-yl ester TFA salt (46 mg,
26%)
[0454] LC/MS=630 (M.sup.++1)
[0455] Retention time: 3.89 min
[0456] Gradient: 0 min-0.2 min 5% ACN, 0.2 min-3.95 min 5%-100%
ACN, 3.95 min-5.20 min 100% ACN, 5.20 min-5.5 min 100%-5% ACN, 5.5
min-6 min 5% ACN.
Example 70
Compound 70--Synthesis of
3-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophene-3-yl]-(trans-4-methyl-
cyclohexanecarbonyl)-3-(S)-amino]-pyrrolidine-1-carboxylic acid
1,1-dioxo-tetrahydro-1.lamda..sup.6-thiophene-3-yl ester
##STR00111##
[0458]
5-(3,3-Dimethyl-but-1-ynyl)-3-[[trans-4-methyl-cyclohexanecarbonyl)-
-pyrrolidin-3-(S)-amino]-thiophene-carboxylic acid (200 mg) was
taken up in ACN (4.0 mL). The heterogeneous solution was allowed to
stir for 5 min, and then DIEA (500 .mu.L, 4.6 mmol) and DMAP (cat)
were added sequentially. The reaction was allowed to stir for 5 min
then
3-chlorocarbonyloxy-1,1-dioxo-tetrahydro-1.lamda..sup.6-thiophene
(.apprxeq.200 mg), which was prepared in a similar manner to method
G, was added in one portion. The reaction was stirred at rt for 15
min and determined to be complete by LC/MS. Solvent was removed
under reduced pressure. The reaction mixture was partitioned
between EtOAc and 2N HCl.sub.(aq). The organics were dried over
Na.sub.2SO.sub.4, solids filtered and solvent removed under reduced
pressure.
3-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophene-3-yl]-(trans-4-methyl-
cyclohexanecarbonyl)-3-(S)-amino]-pyrrolidine-1-carboxylic acid
1,1-dioxo-tetrahydro-1.lamda..sup.6-thiophene-3-yl ester
[0459] TFA salt (37 mg, 19%)
[0460] LC/MS=454.97 (M.sup.+-methylcyclohexylcarbonyl)
[0461] Retention time: 3.56 min
[0462] Gradient: 0 min-0.2 min 5% ACN, 0.2 min-3.95 min 5%-100%
ACN, 3.95 min-5.20 min 100% ACN, 5.20 min-5.5 min 100%-5% ACN, 5.5
min-6 min 5% ACN.
Example 71
Compound
71--4-[[(2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(tr-
ans-4-methyl-cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic
acid 1-tert-butoxycarbonyl-3-methyl-pyrrolidin-3-yl ester
##STR00112##
[0463] And
Example 72
Compound
72--4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(tra-
ns-4-methyl-cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic
acid 3-methyl-pyrrolidin-3-yl ester
##STR00113##
[0465]
5-(3,3-Dimethyl-but-1-ynyl)-3-[(trans-4-methyl-cyclohexanecarbonyl)-
-piperidin-4-yl-amino]-thiophene-2-carboxylic acid (150 mg) was
taken up in ACN (2.0 mL). The heterogeneous solution was allowed to
stir for 5 min, and then DIEA (250 .mu.L, 2.3 mmol) and DMAP (cat)
were added sequentially. The reaction was allowed to stir for 5 min
then 3-chlorocarbonyloxy-3-methylpyrrolidine-carboxylic acid
tert-butyl ester (.apprxeq.200 mg), which was prepared in a similar
manner to method G, was added in one portion. The reaction was
stirred at it for 15 min and determined to be complete by LC/MS.
Solvent was removed under reduced pressure. The reaction mixture
was partitioned between EtOAc and 2N HCl.sub.(aq). The organics
were dried over Na.sub.2SO.sub.4, solids filtered and solvent
removed under reduced pressure.
4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(trans-4-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
1-tert-butoxycarbonyl-3-methyl-pyrrolidin-3-yl ester TFA salt (26
mg, 18%) and
4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(trans--
4-methyl-cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
3-methyl-pyrrolidin-3-yl ester TFA salt (8 mg, 6%) were isolated by
reverse phase HPLC as white solids.
[0466]
4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(trans-4-m-
ethyl-cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
1-tert-butoxycarbonyl-3-methyl-pyrrolidin-3-yl ester
[0467] LC/MS=558.19 (M.sup.+-Boc)
[0468] Retention time: 4.03 min
[0469] Gradient: 0 min-0.2 min 5% ACN, 0.2 min-3.95 min 5%-100%
ACN, 3.95 min-5.20 min 100% ACN, 5.20 min-5.5 min 100%-5% ACN, 5.5
min-6 min 5% ACN.
[0470]
4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(trans-4-m-
ethyl-cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
3-methyl-pyrrolidin-3-yl ester
[0471] LC/MS=558.09 (M.sup.++1)
[0472] Retention time: 3.40 min
[0473] Gradient: 0 min-0.2 min 5% ACN, 0.2 min-3.95 min 5%-100%
ACN, 3.95 min-5.20 min 100% ACN, 5.20 min-5.5 min 100%-5% ACN, 5.5
min-6 min 5% ACN
Example 73
Compound 73--Synthesis of
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(trans-4-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
1-tert-butoxycarbonyl-3-methyl-azetidin-3-yl ester
##STR00114##
[0475]
5-(3,3-Dimethyl-but-1-ynyl)-3-[(4-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid (150 mg) was taken up
in ACN (2.0 mL). The heterogeneous solution was allowed to stir for
5 min, and then DIEA (250 .mu.L, 2.3 mmol) and DMAP (cat) were
added sequentially. The reaction was allowed to stir for 5 min then
3-chlorocarbonyloxy-3-methylpyrrolidine-carboxylic acid tert-butyl
ester (.apprxeq.200 mg), which was prepared in a similar manner to
method G, was added in one portion. The reaction was stirred at rt
for 15 min and determined to be complete by LC/MS. Solvent was
removed under reduced pressure. The reaction mixture was
partitioned between EtOAc and 2N HCl.sub.(aq). The organics were
dried over Na.sub.2SO.sub.4, solids filtered and solvent removed
under reduced pressure.
4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(trans-4-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
1-tert-butoxycarbonyl-3-methyl-azetidin-3-ylester TFA salt (38 mg,
22%) was isolated by reverse phase HPLC as a white solid.
[0476] LC/MS=544.20 (M.sup.+-Boc)
[0477] Retention time: 4.01 min
[0478] Gradient: 0 min-0.2 min 5% ACN, 0.2 min-3.95 min 5%-100%
ACN, 3.95 min-5.20 min 100% ACN, 5.20 min-5.5 min 100%-5% ACN, 5.5
min-6 min 5% ACN.
Example 74
Compound 74--Synthesis of
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(trans-4-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
1,1-dioxo-tetrahydro-1.lamda..sup.6-thiophen-3-yl ester
##STR00115##
[0480]
5-(3,3-Dimethyl-but-1-ynyl)-3-[(trans-4-methyl-cyclohexanecarbonyl)-
-piperidin-4-yl-amino]-thiophene-2-carboxylic acid (200 mg) was
taken up in ACN (4.0 mL). The heterogeneous solution was allowed to
stir for 5 min, then DIEA (350 .mu.L, 2.3 mmol) and DMAP (cat) were
added. The reaction was allowed to stir for 5 min then
3-chlorocarbonyloxy-1,1-dioxo-tetrahydro-1.lamda..sup.6-thiophene
(.apprxeq.200 mg), which was prepared in a similar fashion as
method G, was added in one portion. The reaction was stirred at rt
for 15 min and determined to be complete by LC/MS. Solvent was
removed under reduced pressure. The reaction mixture was
partitioned between EtOAc and 2N HCl.sub.(aq). The organics were
dried over Na.sub.2SO.sub.4, solids filtered and solvent removed
under reduced pressure.
4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(trans-4-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
1,1-dioxo-tetrahydro-1.lamda..sup.6-thiophen-3-yl ester TFA salt
(2.0 mg, 1%) was isolated by reverse phase HPLC as a white
solid.
[0481] LC/MS=469.10 (M.sup.+-methylcyclohexycarbonyl)
[0482] Retention time: 3.70 min
[0483] Gradient: 0 min-0.2 min 5% ACN, 0.2 min-3.95 min 5%-100%
ACN, 3.95 min-5.20 min 100% ACN, 5.20 min-5.5 min 100%-5% ACN, 5.5
min-6 min 5% ACN.
Example 75
Compound 75--Synthesis of
3-{4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(trans-4-meth-
yl-cyclohexanecarbonyl)-cyclohexylcarbamoyloxy}-3-methyl-pyrrolidine-1-car-
boxylic acid tert-butyl ester
##STR00116##
[0485]
3-[4-Amino-cyclohexyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]--
5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid (150 mg)
was taken up in ACN (2.5 mL). The heterogeneous solution was
allowed to stir for 5 min, then DIEA (250 .mu.L, 2.3 mmol) and DMAP
(cat) were added sequentially. The reaction was allowed to stir for
5 min then 3-chlorocarbonyloxy-3-methylpyrrolidine-carboxylic acid
tert-butyl ester (.apprxeq.200 mg), which was prepared in a manner
similar to method G, was added in one portion. The reaction was
stirred at rt for 15 min and determined to be complete by LC/MS.
Solvent was removed under reduced pressure. The reaction mixture
was partitioned between EtOAc and 2N HCl.sub.(aq). The organics
were dried over Na.sub.2SO.sub.4, solids filtered and solvent
removed under reduced pressure.
3-{4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(trans-4-meth-
yl-cyclohexanecarbonyl)-cyclohexylcarbamoyloxy}-3-methyl-pyrrolidine-1-car-
boxylic acid tert-butyl ester TFA salt (84 mg, 47%) was isolated by
reverse phase HPLC as a white solid.
[0486] LC/MS=572.21 (M.sup.+-Boc)
[0487] Retention time: 4.03 min
[0488] Gradient: 0 min-0.2 min 5% ACN, 0.2 min-3.95 min 5%-100%
ACN, 3.95 min-5.20 min 100% ACN, 5.20 min-5.5 min 100%-5% ACN, 5.5
min-6 min 5% ACN.
Example 76
Compound
76--5-(3,3-Dimethyl-but-1-ynyl)-3-{(trans-4-cyclohexanecarbonyl)--
[4-(3-methyl-pyrrolidin-3-yloxycarbonylamino)-cyclohexyl]-amino}-thiophene-
-2-carboxylic acid
##STR00117##
[0490] A mixture of
3-{4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(trans-4-meth-
yl-cyclohexanecarbonyl)-cyclohexylcarbamoyloxy}-3-methyl-pyrrolidine-1-car-
boxylic acid tert-butyl ester TFA salt (50 mg) in CH.sub.2Cl.sub.2
(2.0 mL) and 4N HCl in dioxane (0.5 mL) was stirred at rt for 2 h.
The reaction was determined to be complete by LC/MS. Solvents were
removed under reduced pressure.
5-(3,3-Dimethyl-but-1-ynyl)-3-{(trans-4-cyclohexanecarbonyl)-[4-(3-methyl-
-pyrrolidin-3-yloxycarbonylamino)-cyclohexyl]-amino}-thiophene-2-carboxyli-
c acid TFA salt (32 mg, 73%) was isolated by reverse phase HPLC as
a white solid.
[0491] LC/MS=572.19 (M.sup.++1)
[0492] Retention time: 3.17/3.24 min
[0493] Gradient: 0 min-0.2 min 5% ACN, 0.2 min-3.95 min 5%-100%
ACN, 3.95 min-5.20 min 100% ACN, 5.20 min-5.5 min 100%-5% ACN, 5.5
min-6 min 5% ACN.
Example 77
Compound 77--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-[[4-(1,1-dioxo-tetrahydro-1.lamda..sup.6-th-
iophen-3-yloxycarbonylamino)-cyclohexyl]-(trans-4-methylcyclohexanecarbony-
l)-amino]-thiophene-2-carboxylic acid
##STR00118##
[0495]
3-[4-Amino-cyclohexyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]--
5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid (200 mg)
was taken up in ACN (4.0 mL). The heterogeneous solution was
allowed to stir for 5 min, then DIEA (500 .mu.L, 4.6 mmol) and DMAP
(cat) were added sequentially. The reaction was allowed to stir for
5 min then
3-chlorocarbonyloxy-1,1-dioxo-tetrahydro-1.lamda..sup.6-thiophene
(.apprxeq.200 mg), which was prepared in a similar manner to method
G, was added in one portion. The reaction was stirred at rt for 15
min and determined to be complete by LC/MS. Solvent was removed
under reduced pressure. The reaction mixture was partitioned
between EtOAc and 2N HCl.sub.(aq). The organics were dried over
Na.sub.2SO.sub.4, solids filtered and solvent removed under reduced
pressure.
5-(3,3-Dimethyl-but-1-ynyl)-3-[[4-(1,1-dioxo-tetrahydro-1.lamda..sup.6-th-
iophen-3-yloxycarbonylamino)-cyclohexyl]-(trans-4-methylcyclohexanecarbony-
l)-amino]-thiophene-2-carboxylic acid TFA salt (36 mg, 19%)
[0496] LC/MS=606.86 (M.sup.++1)
[0497] Retention time: 3.61 min
[0498] Gradient: 0 min-0.2 min 5% ACN, 0.2 min-3.95 min 5%-100%
ACN, 3.95 min-5.20 min 100% ACN, 5.20 min-5.5 min 100%-5% ACN, 5.5
min-6 min 5% ACN.
Example 79
Compound 79--Synthesis of
5-(3,3-Dimethyl-but-1-ynyl)-3-{(4-methyl-cyclohexanecarbonyl)-[4-(N'-meth-
yl-N'-pyridin-2-yl-hydrazino)-cyclohexyl]-amino}-thiophene-2-carboxylic
acid
##STR00119##
[0500] 2-Bromopyridine (5 mL, 52.4 mmol) and methylhydrazine (20
mL, 360 mmol) were mixed together in a flask fitted with a
water-condenser. After a few minutes a vigorous exothermic reaction
occurred with refluxing of the methylhydrazine. When the reaction
had subsided, the mixture was left for 16 hours, and the excess
methylhydrazine was removed under reduced pressure. The cooled
residue was stirred with aqueous sodium hydroxide solution (30 ml
of 20%), and the resulting solution was extracted with ether
(3.times.100 ml). The combined extracts were dried
(K.sub.2CO.sub.3) and purified by silica gel chromatography 0-3%
EtOH/CH.sub.2Cl.sub.2 to give 4.8 g of
N-Methyl-N-pyridin-2-yl-hydrazine.
[0501] A mixture of
5-(3,3-Dimethyl-but-1-ynyl)-3-[(4-methyl-cyclohexanecarbonyl)-(4-oxo-cycl-
ohexyl)-amino]-thiophene-2-carboxylic acid methyl ester (412 mg,
0.90 mmol) and N-Methyl-N-pyridin-2-yl-hydrazine (222 mg, 1.8 mmol)
in DCE (6 mL) was treated with AcOH (200 .mu.L, 3.0 mmol) followed
by NaBH(OAc).sub.3 (300 mg, 4.08 mmol) in two or three portions.
After 30 min, NaHCO.sub.3 (saturated aqueous solution, 4-8 mL) was
added to the mixture, followed by brine (20 mL), and the crude
product was extracted with ethyl acetate (2.times.20 mL). The
combined organic layers were concentrated and the crude material
was dissolved in a 3:2:1 mixture of THF:MeOH:water (20 mL), treated
with lithium hydroxide (4.5 mmol, 188 mg) and heated to 60 deg C.
for 2 hours. The residue was purified by HPLC (Gemini column, 35%
acetonitrile:water, 2 min, 35-50% acetonitrile:water, 2 min,
50-100% acetonitrile: water 13 min, both solvents containing 0.1%
trifluoroacetic acid). This resulted in 189 mg (37% yield) of the
title compound as its trans isomer (TFA salt): MS (m/z): 551.3
[M-H].sup.+; HPLC retention time: 3.39 min (2-98%
acetonitrile:water with 0.05% trifluoroacetic acid) and 71 mg (14%
yield) of the title compound as its cis isomer (TFA salt): MS
(m/z): 551.3 [M-H].sup.+; HPLC retention time: 3.52 min (2-98%
acetonitrile:water with 0.05% trifluoroacetic acid).
Example 80
Compound 80--Synthesis of
5-(3,3-Dimethyl-but-1-ynyl)-3-{(4-methyl-cyclohexanecarbonyl)-[4-(N'-meth-
yl-N'-pyrazin-2-yl-hydrazino)-cyclohexyl]-amino}-thiophene-2-carboxylic
acid
##STR00120##
[0503] 2-Chloropyrazine (1 g, 8.3 mmol) and methylhydrazine (1.31
mL, 25 mmol) were mixed together in a flask fitted with a water
condenser. After a few minutes an exotherm was observed. After 16 h
the excess methylhydrazine was removed under reduced pressure. The
cooled residue was stirred with aqueous sodium hydroxide solution
(20 ml of 20%), and the resulting solution was extracted with ether
(6.times.100 ml). The combined extracts were dried
(K.sub.2CO.sub.3) and concentrated to give a light orange solid 370
mg of N-Methyl-N-pyrazin-2-yl-hydrazine.
[0504] A mixture of
5-(3,3-Dimethyl-but-1-ynyl)-3-[(4-methyl-cyclohexanecarbonyl)-(4-oxo-cycl-
ohexyl)-amino]-thiophene-2-carboxylic acid methyl ester (100 mg,
0.22 mmol) and N-Methyl-N-pyrazin-2-yl-hydrazine (41 mg, 0.33 mmol)
in DCE (2 mL) was treated with AcOH (100 .mu.L, 1.5 mmol) followed
by NaBH(OAc).sub.3 (120 mg, 0.5 mmol) in two portions. After 5 h,
NaHCO.sub.3 (saturated aqueous solution, 2 mL) was added to the
mixture, followed by brine (20 mL), and the crude product was
extracted with ethyl acetate (2.times.20 mL). The combined organic
layers were concentrated and the crude material was dissolved in a
3:2:1 mixture of THF:MeOH:water (20 mL), treated with lithium
hydroxide (1.1 mmol, 46 mg) and heated to 60 deg C. for 1 hours.
The residue was purified by HPLC (Gemini column, 35%
acetonitrile:water, 2 min, 35-50% acetonitrile:water, 2 min,
50-100% acetonitrile: water 13 min, both solvents containing 0.1%
trifluoroacetic acid). This resulted in 11 mg of the title compound
as its trans isomer (TFA salt): MS (m/z): 552.2 [M-H].sup.+; HPLC
retention time: 26.05 min (Phenomenex Luna C18, 2-98%
acetonitrile:water with 0.1% trifluoroacetic acid, 30 min gradient)
and 7 mg of the title compound as its cis isomer (TFA salt): MS
(m/z): 552.2 [M-H].sup.+; HPLC retention time: 24.38 min
(Phenomenex Luna C18, 2-98% acetonitrile:water with 0.1%
trifluoroacetic acid, 30 min gradient).
Example 81
Compound 81--Synthesis of
5-(3,3-Dimethyl-but-1-ynyl)-3-{(4-methyl-cyclohexanecarbonyl)-[4-(N'-meth-
yl-N'-thiazol-2-yl-hydrazino)-cyclohexyl]-amino}-thiophene-2-carboxylic
acid
##STR00121##
[0506] 2-Chlorothiazole (0.9 g, 7.52 mmol) was placed in a flask
fitted with a water condenser and methylhydrazine (1.31 mL, 25
mmol) was added dropwise. An immediate exotherm was observed and
methylhydrazine began to reflux. When the reaction had subsided,
the mixture was left for 16 hours, and the excess methylhydrazine
was removed under reduced pressure. The cooled residue was stirred
with aqueous sodium hydroxide solution (20 ml of 20%), and the
resulting solution was extracted with ether (6.times.100 ml). The
combined extracts were dried (K.sub.2CO.sub.3) and concentrated to
give 485 mg of N-Methyl-N-thiazol-2-yl-hydrazine.
[0507] A mixture of
5-(3,3-Dimethyl-but-1-ynyl)-3-[(4-methyl-cyclohexanecarbonyl)-(4-oxo-cycl-
ohexyl)-amino]-thiophene-2-carboxylic acid methyl ester (200 mg,
0.44 mmol) and N-Methyl-N-thiazol-2-yl-hydrazine (112 mg, 0.87
mmol) in DCE (4 mL) was treated with AcOH (200 .mu.L, 3.0 mmol)
followed by NaBH(OAc).sub.3 (120 mg, 0.57 mmol) in two portions.
After 5 h, NaHCO.sub.3 (saturated aqueous solution, 3 mL) was added
to the mixture, followed by brine (20 mL), and the crude product
was extracted with ethyl acetate (2.times.20 mL). The combined
organic layers were concentrated and the crude material was
dissolved in a 3:2:1 mixture of THF:MeOH:water (20 mL), treated
with lithium hydroxide (2.2 mmol, 92 mg) and heated to 60 deg C.
for 1 hours. The residue was purified by HPLC (Gemini column, 35%
acetonitrile:water, 2 min, 35-50% acetonitrile:water, 2 min,
50-100% acetonitrile: water 13 min, both solvents containing 0.1%
trifluoroacetic acid). This resulted in 36 mg of the title compound
as its trans isomer (TFA salt): MS (m/z): 557.2 [M-H].sup.+; HPLC
retention time: 3.43 min (2-98% acetonitrile:water with 0.05%
trifluoroacetic acid) and 8 mg of the title compound as its cis
isomer (TFA salt): MS (m/z): 557.2 [M-H].sup.+; HPLC retention
time: 3.41 min (2-98% acetonitrile:water with 0.05% trifluoroacetic
acid).
Example 82
Compound 82--Synthesis of
5-(3,3-Dimethyl-but-1-ynyl)-3-[{4-[N'-(6-hydroxy-pyridazin-3-yl)-N'-methy-
l-hydrazino]-cyclohexyl}-(4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-
-carboxylic acid
Example 83
Compound
83--[{4-[N'-(6-Chloro-pyridazin-3-yl)-N'-methyl-hydrazino]-cycloh-
exyl}-(4-methyl-cyclohexanecarbonyl)-amino]-5-(3,3-dimethyl-but-1-ynyl)-th-
iophene-2-carboxylic acid
##STR00122##
[0509] 3,6-Dichloro-pyridazine (1 g, 6.75 mmol) was placed in a
flask fitted with a water condenser and methylhydrazine (1.31 mL,
25 mmol) was added dropwise. An immediate exotherm was observed and
methylhydrazine begins to reflux. When the reaction had subsided,
the mixture was left for 16 hours, and the excess methylhydrazine
was removed under reduced pressure. The cooled residue was stirred
with aqueous sodium hydroxide solution (20 ml of 20%), and the
resulting solution was extracted with ether (6.times.100 ml). The
combined extracts were dried (K.sub.2CO.sub.3) and concentrated to
give 427 mg of N-(6-Chloro-pyridazin-3-yl)-N-methyl-hydrazine.
[0510] A mixture of
5-(3,3-Dimethyl-but-1-ynyl)-3-[(4-methyl-cyclohexanecarbonyl)-(4-oxo-cycl-
ohexyl)-amino]-thiophene-2-carboxylic acid methyl ester (500 mg,
1.09 mmol) and N-(6-Chloro-pyridazin-3-yl)-N-methyl-hydrazine (260
mg, 1.64 mmol) in DCE (8 mL) was treated with AcOH (0.5 mL, 8.4
mmol) followed by NaBH(OAc).sub.3 (347 mg, 1.64 mmol) in two
portions. After 5 h, NaHCO.sub.3 (saturated aqueous solution, 10
mL) was added to the mixture, followed by brine (20 mL), and the
crude product was extracted with ethyl acetate (2.times.50 mL). The
combined organic layers were concentrated and the crude material
was dissolved in 4 mL of acetic acid, treated with NaOAc (893 mg,
10.9 mmol) and heated in a sealed tube at 100 deg C. for 16 h. Upon
cooling the mixture was diluted with ethyl acetate and neutralized
with NaHCC.sub.3 (saturated aqueous solution). The combined organic
layers were concentrated and the crude material was dissolved in a
3:2:1 mixture of THF:MeOH:water (20 mL), treated with lithium
hydroxide (5.45 mmol, 228 mg) and heated to 60 deg C. for 1
hour.
[0511] The crude material was purified by HPLC (Gemini column, 35%
acetonitrile:water, 2 min, 35-50% acetonitrile:water, 2 min,
50-100% acetonitrile: water 13 min, both solvents containing 0.1%
trifluoroacetic acid). This resulted in 16 mg of
3-[{4-[N'-(6-Chloro-pyridazin-3-yl)-N'-methyl-hydrazino]-cyclohexyl}-(4-m-
ethyl-cyclohexanecarbonyl)-amino]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2--
carboxylic acid as the trans isomer (TFA salt): MS (m/z): 587.2
[M-H].sup.+; HPLC retention time: 26.39 min (Phenomenex Luna C18,
2-98% acetonitrile:water with 0.1% trifluoroacetic acid, 30 min
gradient); 4 mg of
3-[{4-[N'-(6-Chloro-pyridazin-3-yl)-N'-methyl-hydrazino]-cyclohexyl}-(-
4-methyl-cyclohexanecarbonyl)-amino]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-
-2-carboxylic acid as the cis isomer (TFA salt): MS (m/z): 587.2
[M-H].sup.+; HPLC retention time: 24.03 min (Phenomenex Luna C18,
2-98% acetonitrile:water with 0.1% trifluoroacetic acid, 30 min
gradient); 44 mg of
5-(3,3-Dimethyl-but-1-ynyl)-3-[{4-[N'-(6-hydroxy-pyridazin-3-yl)-N'-
-methyl-hydrazino]-cyclohexyl}-(4-methyl-cyclohexanecarbonyl)-amino]-thiop-
hene-2-carboxylic acid as its trans isomer (TFA salt): MS (m/z):
569.4 [M-H].sup.+; HPLC retention time: 26.15 min (Phenomenex Luna
C18, 2-98% acetonitrile:water with 0.1% trifluoroacetic acid, 30
min gradient), and 71 mg of
5-(3,3-Dimethyl-but-1-ynyl)-3-[{4-[N'-(6-hydroxy-pyridazin-3-yl)-
-N'-methyl-hydrazino]-cyclohexyl}-(4-methyl-cyclohexanecarbonyl)-amino]-th-
iophene-2-carboxylic acid as its cis isomer (TFA salt): MS (m/z):
569.4 [M-H].sup.+; HPLC retention time: 24.67 min (Phenomenex Luna
C18, 2-98% acetonitrile:water with 0.1% trifluoroacetic acid, 30
min gradient).
Hydroxylamine Synthesis
Hydroxylamine 1
Synthesis of O-pyridin-2-ylmethylhydroxylamine dihydrochloride
##STR00123##
[0513] 2-Chloromethylpyridine hydrochloride (514 mg, 3.13 mmol),
N-hydroxyphthalimide (515 mg, 3.18 mmol), and triethylamine (1.3
mL, 9.3 mmol) were mixed in acetonitrile (5 mL) and stirred at
80.degree. C. for 2.5 hours. The reaction mixture was diluted with
ethyl acetate and washed sequentially with 1N NaOH.sub.(aq), water,
and brine. The organic phase was dried over MgSO.sub.4, filtered,
and concentrated. The residue was recrystallized from ethyl acetate
and hexanes to provide N-(pyridine-2-ylmethoxy)phthalimide (364 mg,
1.44 mmol).
[0514] Hydrazine monohydrate (53 .mu.L, 1.5 mmol) was added to a
solution of N-(pyridine-2-ylmethoxy)phthalimide (356 mg; 1.41 mmol)
in ethanol (2 mL) and stirred at 80.degree. C. for one hour in a
sealed tube. The reaction mixture was diluted with ethyl acetate,
adsorbed onto silica gel, and purified by SGC (0-20% [8:1
EtOH:NH.sub.4OH.sub.(aq)]/DCM). After concentration of fractions
the residue was taken up in diethyl ether and treated with 4N
HCl.sub.(Et2O) to provide O-pyridin-2-ylmethylhydroxylamine
dihydrochloride (152 mg, 0.77 mmol) as a white solid.
[0515] Using the same procedure as described for
O-pyridin-2-ylmethylhydroxylamine dihydrochloride, the
hydroylamines shown below were syntheisized.
##STR00124##
Example 112
Compound 112--Synthesis of
5-(3,3-Dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-[4-(p-
yridin-2-ylmethoxyimino)-cyclohexyl]-amino}-thiophene-2-carboxylic
acid
##STR00125##
[0517]
5-(3,3-dimethyl-but-1-ynyl)-3-[(trans-4-methyl-cyclohexanecarbonyl)-
-(4-oxo-cyclohexyl)-amino]-thiophene-2-carboxylic acid methyl ester
(264 mg, 0.58 mmol), O-pyridin-2-ylmethylhydroxylamine
dihydrochloride (142 mg, 0.72 mmol), and sodium acetate (177 mg,
2.2 mmol) were mixed in 2:1 MeOH:DCM (3 mL) and stirred at
50.degree. C. for 1 hour. Lithium hydroxide monohydrate (240 mg,
5.7 mmol) and water (1 mL) were then added and the reaction mixture
continued to stir at 50.degree. C. for 2 hours. The reaction
mixture was then partitioned between ethyl acetate and 5% citric
acid.sub.(aq). The aqueous phase was neutralized with 4N
NaOH.sub.(aq) and thrice extracted with ethyl acetate. The combined
organic phases were washed with brine, dried over MgSO.sub.4,
filtered, and concentrated. The residue was purified by HPLC
(Gemini column; 25% acetonitrile:water, 2 min; 25-100%
acetonitrile:water, 16 min; 100% acetonitrile, 3 min; both solvents
containing 0.1% trifluoroacetic acid). This resulted in 234 mg (19%
yield over 2 steps) of the title compound as a white powder (TFA
salt): MS (m/z): 550.2 [M+H].sup.+; HPLC retention time: 3.55 min
(2-98% acetonitrile:water with 0.05% trifluoroacetic acid).
Example 113
Compound
113--3-(N-(4-Benzyloxyimino)cyclohexyl)-4-methylcyclohexanecarbox-
amido)-5-(3,3-dimethylbut-1-ynyl)thiophene-2-carboxylic acid
##STR00126##
[0519] Prepared in the same manner as Example 112 using
O-benzylhydroxylamine. MS (m/z): 549.1 [M+H].sup.+; HPLC retention
time: 5.23 min.
Example 114
Compound
114--3-(N-(4-((1H-1,2,4-Triazol-1-yl)methoxyimino)cyclohexyl)-4-m-
ethylcyclohexanecarboxamido)-5-(3,3-dimethylbut-1-ynyl)thiophene-2-carboxy-
lic acid
##STR00127##
[0521] Prepared in the same manner as Example 112 using
hydroxylamine 2. MS (m/z): 540.1 [M+H].sup.+; HPLC retention time:
4.29 min.
Example 115
Compound
115--3-(N-(4-(Carboxymethoxyimino)cyclohexyl)-4-methylcyclohexane-
carboxamido)-5-(3,3-dimethylbut-1-ynyl)thiophene-2-carboxylic
acid
##STR00128##
[0523] Prepared in the same manner as Example 112 using
hydroxylamine 5. MS (m/z): 517.0 [M+H].sup.+; HPLC retention time:
4.28 min.
Example 116
Compound
116--3-(N-(4-(Cyanomethoxyimino)cyclohexyl)-4-methylcyclohexaneca-
rboxamido)-5-(3,3-dimethylbut-1-ynyl)thiophene-2-carboxylic
acid
##STR00129##
[0525]
5-(3,3-Dimethyl-but-1-ynyl)-3-[(trans-4-methyl-cyclohexanecarbonyl)-
-(4-oxo-cyclohexyl)-amino]-thiophene-2-carboxylic acid methyl ester
(519 mg, 1.13 mmol) was dissolved in a mixture of 3:2:1
THF:MeOH:H.sub.2O (25 mL). Lithium hydroxide (5 mL, 1.0N aqueous
solution) was added and the reaction mixture was stirred at
60.degree. C. for 1 hour. The reaction mixture was then partitioned
between ethyl acetate and water. The aqueous phase was neutralized
with 5% citric acid.sub.(aq), and thrice extracted with ethyl
acetate. The combined organic phases were washed with brine, dried
over MgSO.sub.4, filtered, and concentrated to afford 505 mg of
5-(3,3-dimethyl-but-1-ynyl)-3-[(trans-4-methyl-cyclohexanecarbonyl)-(4-ox-
o-cyclohexyl)-amino]-thiophene-2-carboxylic acid which was carried
on without further purification.
[0526]
5-(3,3-Dimethyl-but-1-ynyl)-3-[(trans-4-methyl-cyclohexanecarbonyl)-
-(4-oxo-cyclohexyl)-amino]-thiophene-2-carboxylic acid (505 mg,
1.02 mmol), O-cyanomethylhydroxylamine hydrochloride (174 mg, 1.32
mmol), and sodium acetate (212 mg, 2.6 mmol) were mixed in 2:1
MeOH:DCM (7.5 mL) and stirred at 50.degree. C. for 1 hour. The
reaction mixture was then partitioned between ethyl acetate and
water. The aqueous phase was thrice extracted with ethyl acetate
and the combined organic layers were washed with brine, dried over
MgSO.sub.4, filtered, and concentrated. The residue was purified by
HPLC (Gemini column; 25% acetonitrile:water, 2 min; 25-100%
acetonitrile:water, 16 min; 100% acetonitrile, 3 min; both solvents
containing 0.1% trifluoroacetic acid). This resulted in 159 mg (28%
yield over 2 steps) of the title compound as a white powder: MS
(m/z): 495.7 [M-H].sup.-; HPLC retention time: 4.68 min (2-98%
acetonitrile:water with 0.05% trifluoroacetic acid).
Example 117
Compound
117--5-(3,3-Dimethylbut-1-ynyl)-3-(4-methyl-N-(4-(pyridin-4-ylmet-
hoxyimino)cyclohexyl)cyclohexanecarboxamido)thiophene-2-carboxylic
acid
##STR00130##
[0528] Prepared in the same manner as Example 112 using
hydroxylamine 3. MS (m/z): 550.1 [M+H].sup.+; HPLC retention time:
3.46 min.
Example 118
Compound
118--5-(3,3-Dimethylbut-1-ynyl)-3-(4-methyl-N-(4-(pyridin-3-ylmet-
hoxyimino)cyclohexyl)cyclohexanecarboxamido)thiophene-2-carboxylic
acid
##STR00131##
[0530] Prepared in the same manner as Example 112 using
hydroxylamine 4. MS (m/z): 550.1 [M+H].sup.+; HPLC retention time:
3.47 min.
Example 119
Compound
119--5-(3,3-Dimethylbut-1-ynyl)-3-(4-methyl-N-(4-(phenoxyimino)cy-
clohexyl)cyclohexanecarboxamido)thiophene-2-carboxylic acid
##STR00132##
[0532] Prepared in the same manner as Example 112 using
O-phenylhydroxylamine. MS (m/z): 535.0 [M+H].sup.+; HPLC retention
time: 5.38 min.
Example 120
Compound
120--5-(3,3-Dimethylbut-1-ynyl)-3-(N-(4-(methoxyimino)cyclohexyl)-
-4-methylcyclohexanecarboxamido)thiophene-2-carboxylic acid
##STR00133##
[0534] Prepared in the same manner as Example 112 using
O-methylhydroxylamine. MS (m/z): 473.0 [M+H].sup.+; HPLC retention
time: 4.79 min.
Example 121
Compound
121--3-(N-(4-(4-(Methylsulfonyl)benzyloxyimino)cyclohexyl)-4-meth-
ylcyclohexanecarboxamido)-5-(3,3-dimethylbut-1-ynyl)thiophene-2-carboxylic
acid
##STR00134##
[0536] Prepared in the same manner as Example 112 using
hydroxylamine 6. MS (m/z): 627.0 [M+H].sup.+; HPLC retention time:
4.75 min.
Example 122
Compound 122--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-[(N'-(1,4-dioxa-spiro[4.5]dec-8-yl)-N'-meth-
yl-N-(4-methyl-cyclohexanecarbonyl)-hydrazino]-thiophene-2-carboxylic
acid
##STR00135##
[0538] To a solution of
3-tert-butoxycarbonylamino-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carbox-
ylic acid methyl ester (4.3 g, 12.8 mmol) in NMP (64 mL) was added
K-OtBu (1.6 g, 14.1 mmol). After 10 min a solution of
0-4-nitrobenzoylhydroxylamine (3.0 g, 16.6 mmol) in NMP (16 mL) was
added dropwise and the reaction was allowed to stir for 16 h. The
reaction was quenched with LiCl solution (5% in water, 50 mL) and
diluted with EtOAc (50 mL). The crude product was washed 3.times.50
mL with 5% LiCl solution, dried over Na.sub.2SO.sub.4, and purified
by silica gel chromatography 0-30% EtOAc/hexanes to give
3-(N-tert-butoxycarbonyl-hydrazino)-5-(3,3-dimethyl-but-1-ynyl)-thiophene-
-2-carboxylic acid methyl ester (2.6 g, 7.4 mmol).
[0539]
3-(N-tert-Butoxycarbonyl-hydrazino)-5-(3,3-dimethyl-but-1-ynyl)-thi-
ophene-2-carboxylic acid methyl ester (2.6 g, 7.4 mmol) in
Et.sub.2O (37 mL) was cooled to 0.degree. C. and treated with
trifluoroacetic anhydride (1.9 g, 8.9 mmol) for 1 h. The reaction
was concentrated, dissolved with neat TFA (4.3 g, 37 mmol) and
heated to 60.degree. C. After 15 min, the reaction was cooled to
rt, diluted with toluene (30 mL) and concentrated, this step was
repeated three times, to give crystalline
5-(3,3-Dimethyl-but-1-ynyl)-3-[N'-(2,2,2-trifluoro-acetyl)-hydrazino]-thi-
ophene-2-carboxylic acid methyl ester (2.1 g, 6.0 mmol).
[0540] A mixture of
5-(3,3-dimethyl-but-1-ynyl)-3-[N'-(2,2,2-trifluoro-acetyl)-hydrazino]-thi-
ophene-2-carboxylic acid methyl ester (2.1 g, 6.0 mmol),
4-methyl-cyclohexanecarbonyl chloride (1.45 g, 9 mmol), DMAP (1.1
g, 9 mmol) and DCE (20 mL) was heated to 40.degree. C. for 16 h.
After cooling, the mixture was concentrated and the product was
purified by silica gel chromatography 0-30% EtOAc/hexanes to give
5-(3,3-dimethyl-but-1-ynyl)-3-[N-(4-methyl-cyclohexanecarbonyl)-N'-(2,2,2-
-trifluoro-acetyl)-hydrazino]-thiophene-2-carboxylic acid methyl
ester (2.74 g, 5.8 mmol).
[0541] To a solution of
5-(3,3-dimethyl-but-1-ynyl)-3-[N-(4-methyl-cyclohexanecarbonyl)-N'-(2,2,2-
-trifluoro-acetyl)-hydrazino]-thiophene-2-carboxylic acid methyl
ester (1.6 g, 3.4 mmol) in THF (17 mL) was added K-OtBu (4.1 mmol,
1.0 M in THF), followed by MeI (1.0 g, 6.8 mmol). After stirring at
rt for 16 h the reaction was diluted with EtOAc washed once with 20
mL of brine and purified by silica gel chromatography 0-20%
EtOAc/hexanes to give
5-(3,3-dimethyl-but-1-ynyl)-3-[N'-methyl-N-(4-methyl-cyclohexanecarbonyl)-
-N'-(2,2,2-trifluoro-acetyl)-hydrazino]-thiophene-2-carboxylic acid
methyl ester. This was then diluted with EtOAc (20 mL) and treated
with NaOH (6.8 mL of 2M aq solution) to give
5-(3,3-Dimethyl-but-1-ynyl)-3-[N'-methyl-N-(4-methyl-cyclohexanecarbonyl)-
-hydrazino]-thiophene-2-carboxylic acid.
[0542] A mixture of
5-(3,3-Dimethyl-but-1-ynyl)-3-[N'-methyl-N-(4-methyl-cyclohexanecarbonyl)-
-hydrazino]-thiophene-2-carboxylic acid (300 mg, 0.8 mmol),
1,4-cyclohexadione-mono-ethylene ketal (149 mg, 1 mmol), AcOH (300
mg, 5 mmol) in DCE (2 mL) was treated with NaBH(OAc).sub.3 (430 mg,
2 mmol) for 16 h. The reaction was quenched with water (10 mL) and
extracted with EtOAc. A portion of this material was then purified
by HPLC (Gemini column, 35% acetonitrile:water, 2 min, 35-50%
acetonitrile:water, 2 min, 50-100% acetonitrile: water 13 min, both
solvents containing 0.1% trifluoroacetic acid) giving
5-(3,3-dimethyl-but-1-ynyl)-3-[N'-(1,4-dioxa-spiro[4.5]dec-8-yl)-N'-methy-
l-N-(4-methyl-cyclohexanecarbonyl)-hydrazino]-thiophene-2-carboxylic
acid as its TFA salt: MS (m/z): 517.1 [M-H].sup.+; HPLC retention
time: 4.73 min (2-98% acetonitrile:water with 0.05% trifluoroacetic
acid).
Example 123 (Compound 123) and 124 (Compound 124)
Synthesis of
3-(2-(4-hydroxycyclohexyl)-2-methyl-1-(4-methylcyclohexanecarbonyl)hydraz-
inyl)-5-(3,3-dimethylbut-1-ynyl)thiophene-2-carboxylic acid (123)
and
3-(2-(4-hydroxycyclohexyl)-2-methyl-1-(4-methylcyclohexanecarbonyl)hydraz-
inyl)-5-(3,3-dimethylbut-1-ynyl)thiophene-2-carboxylic acid
(124)
##STR00136##
[0544] A mixture of
5-(3,3-dimethyl-but-1-ynyl)-3-[N'-(1,4-dioxa-spiro[4.5]dec-8-yl)-N'-methy-
l-N-(4-methyl-cyclohexanecarbonyl)-hydrazino]-thiophene-2-carboxylic
(500 mg, 1 mmol) in a 1:1 mixture of THF and MeOH (2.5 mL) was
treated with 1 M HCl (2.5 mL) and heated for 3 h at 60.degree. C.
The reaction mixture was cooled to room temperature, diluted with
EtOAc (10 mL), neutralized with sat. NaHCO.sub.3, washed once with
brine (10 mL), dried over Na.sub.2SO.sub.4, and concentrated. The
crude material was dissolved in wet THF (5 mL) and treated with
NaBH.sub.4 (38 mg, 1.1 mmol) at 0.degree. C. for 30 min. A portion
of
5-(3,3-dimethyl-but-1-ynyl)-3-[N'-(4-hydroxy-cyclohexyl)-N'-methyl-N-(4-m-
ethyl-cyclohexanecarbonyl)-hydrazino]-thiophene-2-carboxylic acid
was purified by HPLC (Gemini column, 35% acetonitrile:water, 2 min,
35-50% acetonitrile:water, 2 min, 50-100% acetonitrile: water 13
min, both solvents containing 0.1% trifluoroacetic acid) to give 20
mg as its TFA salt: MS (m/z): 475.7 [M-H].sup.+; HPLC retention
time: 4.33 min (2-98% acetonitrile:water with 0.05% trifluoroacetic
acid).
[0545] Additional HPLC purification separated Example 123
(retention time: 27.89 min) from Example 124 (retention time
27.89).
Example 125 and 126
Synthesis of
5-(3,3-dimethylbut-1-ynyl)-3-(2-methyl-1-(4-methylcyclohexanecarbonyl)-2--
(4-(pyridin-3-yloxy)cyclohexyl)hydrazinyl)thiophene-2-carboxylic
acid (Compound 125) and
5-(3,3-dimethylbut-1-ynyl)-3-(2-methyl-1-(4-methylcyclohexanecarbonyl)-2--
(4-(pyridin-3-yloxy)cyclohexyl)hydrazinyl)thiophene-2-carboxylic
acid (Compound 126)
##STR00137##
[0547] A mixture of
5-(3,3-dimethyl-but-1-ynyl)-3-[N'-(4-hydroxy-cyclohexyl)-N'-methyl-N-(4-m-
ethyl-cyclohexanecarbonyl)-hydrazino]-thiophene-2-carboxylic acid
(100 mg, 0.2 mmol) and 3-fluoro-pyridine (97 .mu.L, 1 mmol) in DMF
(0.6 mL) was treated with sodium hydride (40 mg, 1 mmol, 60% oil
dispersion) in two or three portions. The mixture was stirred until
the bubbling slowed, and was sealed and heated at 100 deg C. for 8
h. After cooling, ethyl acetate (2-3 mL) was added and the mixture
was carefully quenched with citric acid (10% aqueous solution, 2-3
mL). Water was added and the mixture was extracted with ethyl
acetate (2.times.30 mL). The combined organic layers were dried
over sodium sulfate, filtered and concentrated. The residue was
purified by HPLC (Gemini column, 35% acetonitrile:water, 2 min,
35-50% acetonitrile:water, 2 min, 50-95% acetonitrile: water 13
min, both solvents containing 0.1% trifluoroacetic acid). This
resulted in 37 mg of the title compound as a white powder (bis-TFA
salt): MS (m/z): 552.1 [M-H].sup.+; HPLC retention time: 3.50 min
(2-98% acetonitrile:water with 0.05% trifluoroacetic acid).
[0548] Additional HPLC purification separated Example 125
(retention time: 22.46 min) from Example 126 (retention time
22.62).
Example 127
Compound 127--Synthesis of
4-[N'-[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-N-methyl-N'-(-
4-methyl-cyclohexanecarbonyl)-hydrazino]-piperidine-1-carboxylic
acid tert-butyl ester
##STR00138##
[0550] The title compound was synthesized in a manner analogous to
Example 122, using 1-N-Boc-4-piperidone in place of
1,4-cyclohexadione-mono-ethylene ketal: MS (m/z): 560.1
[M-H].sup.+; HPLC retention time: 5.10 min (2-98%
acetonitrile:water with 0.05% trifluoroacetic acid).
Example 128
Compound 128--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-[N'-methyl-N-(4-methyl-cyclohexanecarbonyl)-
-N'-piperidin-4-yl-hydrazino]-thiophene-2-carboxylic acid
##STR00139##
[0552] Crude
4-[N'-[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-N-methyl-N'-(-
4-methyl-cyclohexanecarbonyl)-hydrazino]-piperidine-1-carboxylic
acid tert-butyl ester (220 mg, 500 mmol) (Example 127) was treated
with TFA (3 mL, 4.6 mmol) at 60.degree. C. for 10 min. After
cooling, toluene (2-3 mL) was added and the mixture was
concentrated, this was repeated several times, and a portion of
this crude material was purified by HPLC (Gemini column, 35%
acetonitrile:water, 2 min, 35-50% acetonitrile:water, 2 min, 50-95%
acetonitrile: water 13 min, both solvents containing 0.1%
trifluoroacetic acid). This resulted in 20 mg of the title compound
as a white powder (bis-TFA salt): MS (m/z): 460.2 [M-H].sup.+; HPLC
retention time: 3.15 min (2-98% acetonitrile:water with 0.05%
trifluoroacetic acid).
Example 129
Compound 129--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-[N'-methyl-N-(4-methyl-cyclohexanecarbonyl)-
-N'-(1-methyl-piperidin-4-yl)-hydrazino]-thiophene-2-carboxylic
acid
##STR00140##
[0554] A mixture of
5-(3,3-dimethyl-but-1-ynyl)-3-[N'-methyl-N-(4-methyl-cyclo-hexanecarbonyl-
)-N'-piperidin-4-yl-hydrazino]-thiophene-2-carboxylic acid (100 mg,
225 mmol) (Example 128), formaldehyde (1.12 mmol), acetic acid (0.5
mL, 8 mmol) in DCE (3 mL) was treated with NaBH(OAc).sub.3 (36 mg,
168 mmol) for 16 h. The reaction was quenched with water (10 mL)
and extracted with EtOAc. The crude material was then purified by
HPLC (Gemini column, 35% acetonitrile:water, 2 min, 35-50%
acetonitrile:water, 2 min, 50-100% acetonitrile: water 13 min, both
solvents containing 0.1% trifluoroacetic acid) giving
5-(3,3-dimethyl-but-1-ynyl)-3-[N'-methyl-N-(4-methyl-cyclohexanecarbonyl)-
-N'-(1-methyl-piperidin-4-yl)-hydrazino]-thiophene-2-carboxylic
acid as its bis-TFA salt: MS (m/z): 474.2 [M-H].sup.+; HPLC
retention time: 3.17 min (2-98% acetonitrile:water with 0.05%
trifluoroacetic acid).
Example 130
Compound 130--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-{N'-methyl-N-(4-methyl-cyclohexanecarbonyl)-
-N'-[2-(pyridin-3-yloxy)-ethyl]-hydrazino}-thiophene-2-carboxylic
acid
##STR00141##
[0556] The title compound was synthesized in a manner analogous to
Example 122, using (pyridin-3-yloxy)-acetaldehyde in place of
1,4-cyclohexadione-mono-ethylene ketal: MS (m/z): 498.1
[M-H].sup.+; HPLC retention time: 3.31 min (2-98%
acetonitrile:water with 0.05% trifluoroacetic acid).
Example 131
Compound 131--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-[N'-methyl-N-(4-methyl-cyclohexanecarbonyl)-
-N'-(tetrahydro-pyran-4-ylmethyl)-hydrazino]-thiophene-2-carboxylic
acid
##STR00142##
[0558] To a solution of
5-(3,3-dimethyl-but-1-ynyl)-3-[N-(4-methyl-cyclohexanecarbonyl)-N'-(2,2,2-
-trifluoro-acetyl)-hydrazino]-thiophene-2-carboxylic acid methyl
ester (1.6 g, 3.4 mmol) in EtOAc (20 mL) was added NaOH (6.8 mL of
2M aq solution). After 2 h stirring at rt, the reaction was
neutralized with 1M HCl aq, extracted with EtOAc, dried over
Na.sub.2SO.sub.4, and concentrated to give
5-(3,3-dimethyl-but-1-ynyl)-3-[N-(4-methyl-cyclohexanecarbonyl)-hydrazino-
]-thiophene-2-carboxylic acid (1.1 g, 3 mmol) as an off white
solid.
[0559] A mixture of
5-(3,3-dimethyl-but-1-ynyl)-3-[N-(4-methyl-cyclohexanecarbonyl)-hydrazino-
]-thiophene-2-carboxylic acid (50 mg, 0.132 mmol),
tetrahydro-pyran-4-carbaldehyde (19 mg, 0.172 mmol), AcOH (24 mg,
0.4 mmol) in DCE (2 mL) was treated with NaBH(OAc).sub.3 (42 mg,
0.2 mmol) for 16 h. The reaction was quenched with water (10 mL),
extracted with EtOAc and concentrated. A mixture of the crude
material, AcOH (47 mg, 2.37 mmol), dissolved in iPrOH (3 mL) was
treated with NaCNBH.sub.3 (13 mg, 0.198 mmol) then heated to
50.degree. C. for 16 h. The product was then purified by HPLC
(Gemini column, 35% acetonitrile:water, 2 min, 35-50%
acetonitrile:water, 2 min, 50-100% acetonitrile: water 13 min, both
solvents containing 0.1% trifluoroacetic acid) giving
5-(3,3-dimethyl-but-1-ynyl)-3-{N'-methyl-N-(4-methyl-cyclohexanecarbonyl)-
-N'-(tetrahydro-pyran-4-ylmethyl)-hydrazino}-thiophene-2-carboxylic
acid as its TFA salt: MS (m/z): 475.1 [M-H].sup.+; HPLC retention
time: 4.62 min (2-98% acetonitrile:water with 0.05% trifluoroacetic
acid).
Example 132
Compound 132--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-{N'-methyl-N-(4-methyl-cyclohexanecarbonyl)-
-N'-[2-(pyridin-2-yloxy)-ethyl]-hydrazino}-thiophene-2-carboxylic
acid
##STR00143##
[0561] The title compound was synthesized in a manner analogous to
Example 131, using (pyridin-2-yloxy)-acetaldehyde in place of
tetrahydro-pyran-4-carbaldehyde: MS (m/z): 498.1 [M-H].sup.+; HPLC
retention time: 29.8 min (2-98% acetonitrile:water with 0.05%
trifluoroacetic acid) 30 min run.
Example 133
Compound 133--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-{N-(4-methyl-cyclohexanecarbonyl)-N'-[2-(py-
ridin-3-yloxy)-ethyl]-hydrazino}-thiophene-2-carboxylic acid
##STR00144##
[0563] The title compound was synthesized from
5-(3,3-dimethyl-but-1-ynyl)-3-[N-(4-methyl-cyclohexanecarbonyl)-hydrazino-
]-thiophene-2-carboxylic acid and (pyridin-3-yloxy)-acetaldehyde in
a manner similar to that of Example 131. MS (m/z): 484.0
[M-H].sup.+; HPLC retention time: 3.44 min (2-98%
acetonitrile:water with 0.05% trifluoroacetic acid).
Example 134
Compound 134--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-[N'-(2-hydroxy-ethyl)-N-(4-methyl-cyclohexa-
necarbonyl)-N'-oxetan-3-yl-hydrazino]-thiophene-2-carboxylic
acid
##STR00145##
[0565] The title compound was synthesized in a manner analogous to
Example 131, using 3-(tert-butyl-dimethyl-silanyloxy)-acetaldehyde
in place of tetrahydro-pyran-4-carbaldehyde, treating the crude
with TBAF (1 mmol, 0.1 M in THF) prior to the second reductive
amination in which formaldehyde is replaced with oxetan-3-one: MS
(m/z): 461.2 [M-H].sup.-; HPLC retention time: 4.38 min (2-98%
acetonitrile:water with 0.05% trifluoroacetic acid).
Example 135
Compound 135--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-[N'-(2-hydroxy-ethyl)-N'-methyl-N-(4-methyl-
-cyclohexanecarbonyl)-hydrazino]-thiophene-2-carboxylic acid
##STR00146##
[0567] The title compound was synthesized in a manner analogous to
Example 131, using 3-(tert-butyl-dimethyl-silanyloxy)-acetaldehyde
in place of tetrahydro-pyran-4-carbaldehyde and treating the crude
with TBAF (1 mmol, 0.1 M in THF) prior to the second reductive
amination with formaldehyde: MS (m/z): 422.1 [M-H].sup.+; HPLC
retention time: 4.06 min (2-98% acetonitrile:water with 0.05%
trifluoroacetic acid).
Example 136
Compound 136--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-[N'-dimethyl-N-(4-methyl-cyclohexanecarbony-
l)-hydrazino]-thiophene-2-carboxylic acid
##STR00147##
[0569] A mixture of
5-(3,3-dimethyl-but-1-ynyl)-3-iodo-thiophene-2-carboxylic acid
methyl ester (0.200 g, 0.574 mmol), LiCl (0.048, 1.14 mmol),
N,N-dimethylhydrazine (0.043 mL, 0.57 mmol), Pd.sub.2(dba).sub.3
(0.003 g, 0.005 mmol), xantphos (0.008 g, 0.005 mmol), and NaOtBu
(0.037 g, 0.394 mmol) in toluene (3 mL) was heated to 80.degree. C.
for 16 h. The reaction was diluted with ethyl acetate filtered
through a Celite pad and concentrated the crude material was
purified by silica gel chromatography (0-15% EtOAc/hexane) to give
5-(3,3-dimethyl-but-1-ynyl)-3-(N'-dimethyl-hydrazino)-thiophene-2-carboxy-
lic acid methyl ester in 63% yield. This was then dissolved in
pyridine (5 mL) and treated with neat
trans-4-methyl-cyclohexanecarbonyl chloride (0.132 g, 0.825 mmol).
The reaction was heated for 16 h at 85.degree. C. and quenched with
saturated ammonium chloride solution. The reaction mixture was
diluted with EtOAc, washed with water, brine, dried over sodium
sulfate, filtered and concentrated. The crude material was then
dissolved in a 3:2:1 mixture of THF:MeOH:water (5 mL), treated with
lithium hydroxide monohydrate (0.69 g, 1.65 mmol) and heated to
60.degree. C. for 1 hour. The organic volatiles were evaporated
under reduced pressure and the crude material was purified by HPLC
to afford the title compound. MS (m/z): 456.0 [M+H].sup.-; HPLC
retention time: 3.91 min (2-98% acetonitrile:water with 0.05%
trifluoroacetic acid).
Example 137
Compound 137--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-[N'-methyl-N-(4-methyl-cyclohexanecarbonyl)-
-N'-pyridin-2-yl-hydrazino]-thiophene-2-carboxylic acid
##STR00148##
[0571] A mixture of
5-(3,3-dimethyl-but-1-ynyl)-3-iodo-thiophene-2-carboxylic acid
methyl ester (0.100 g, 0.28 mmol), N,N-diethylenediamine (9 .mu.L,
0.086 mmol), N-methyl-N-pyridin-2-yl-hydrazine (0.105 g, 0.86
mmol), CuI (0.01 g, 0.057 mmol), 4 .ANG. ms (0.114 mg) and
K.sub.2CO.sub.3 (0.118 g, 0.861 mmol) in DMF (3 mL) was heated to
80.degree. C. for 16 h. The reaction was diluted with ethyl
acetate, washed twice with 5% LiCl, concentrated and the crude
material was purified by silica gel chromatography (0-20% EtOH in
DCM) to give
5-(3,3-dimethyl-but-1-ynyl)-3-(N'-methyl-N'-pyridin-2-yl-hydrazino)-thiop-
hene-2-carboxylic acid methyl ester in 50% yield. This was then
dissolved in pyridine (5 mL) and treated with neat
trans-4-methyl-cyclohexanecarbonyl chloride (0.132 g, 0.825 mmol).
The reaction was heated for 16 h at 85.degree. C. and quenched with
saturated ammonium chloride solution. The reaction mixture was
diluted with EtOAc, washed with water, brine, dried over sodium
sulfate, filtered and concentrated. The crude material was then
dissolved in a 3:2:1 mixture of THF:MeOH:water (5 mL), treated with
lithium hydroxide monohydrate (0.69 g, 1.65 mmol) and heated to
60.degree. C. for 1 hour. The organic volatiles were evaporated
under reduced pressure and the crude material was purified by HPLC
to afford the title compound. MS (m/z): 454.1 [M+H].sup.-; HPLC
retention time: 3.95 min (2-98% acetonitrile:water with 0.05%
trifluoroacetic acid).
Example 140
Compound 140--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-{(4-methyl-cyclohexanecarbonyl)-[1-(pyridin-
e-3-carbonyl)-piperidin-4-yl]-amino}-thiophene-2-carboxylic
acid
##STR00149##
[0573] A mixture of
5-(3,3-Dimethyl-but-1-ynyl)-3-[(4-methyl-cyclohexanecarbonyl)-piperidin-4-
-yl-amino]-thiophene-2-carboxylic acid methyl ester (63 mg, 0.141
mmol) and pyridine-2-carbonyl chloride HCl salt (33 mg, 0.184 mmol)
in DCM (2 mL) was treated with DIEA (124 .mu.L, 0.7 mmol). After 30
min, NaHCO.sub.3 (saturated aqueous solution, 4-8 mL) was added to
the mixture, followed by brine (20 mL), and the crude product was
extracted with ethyl acetate (2.times.20 mL). The combined organic
layers were concentrated and the crude material was dissolved in a
3:2:1 mixture of THF:MeOH:water (5 mL), treated with lithium
hydroxide (42 mg, 1 mmol,) and heated to 60 deg C. for 2 hours. The
residue was purified by HPLC (Gemini column, 35%
acetonitrile:water, 2 min, 35-50% acetonitrile:water, 2 min,
50-100% acetonitrile: water 13 min, both solvents containing 0.1%
trifluoroacetic acid). This resulted in 40 mg of the title compound
as its TFA salt: MS (m/z): 536.9 [M-H].sup.+; HPLC retention time:
4.17 min (2-98% acetonitrile:water with 0.05% trifluoroacetic
acid).
Example 141
Compound 141--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-{(4-methyl-cyclohexanecarbonyl)-[1'-(pyrazi-
ne-2-carbonyl)-piperidin-4-yl]-amino}-thiophene-2-carboxylic
acid
##STR00150##
[0575] The title compound was synthesized in a manner analogous to
Example 140, using pyrazine-2-carbonyl chloride in place of
pyridine-2-carbonyl chloride HCl salt: MS (m/z): 538.0 [M-H].sup.+;
HPLC retention time: 3.52 min (2-98% acetonitrile:water with 0.05%
trifluoroacetic acid) 30 min run.
Example 142
Compound 142--Synthesis of
5-(3,3-Dimethyl-but-1-ynyl)-3-{(4-methyl-cyclohexanecarbonyl)-[1-(pyridin-
e-3-carbonyl)-pyrrolidin-3-yl]-amino}-thiophene-2-carboxylic
acid
##STR00151##
[0577] A mixture of
5-(3,3-Dimethyl-but-1-ynyl)-3-[(4-methyl-cyclohexanecarbonyl)-pyrrolidin--
3-yl-amino]-thiophene-2-carboxylic acid methyl ester (74 mg, 0.158
mmol) and pyridine-2-carbonyl chloride HCl salt (56 mg, 0.31 mmol)
in DCM (2 mL) was treated with DIEA (124 .mu.L, 0.8 mmol). After 30
min, NaHCO.sub.3 (saturated aqueous solution, 4-8 mL) was added to
the mixture, followed by brine (20 mL), and the crude product was
extracted with ethyl acetate (2.times.20 mL). The combined organic
layers were concentrated and the crude material was dissolved in a
3:2:1 mixture of THF:MeOH:water (5 mL), treated with lithium
hydroxide (42 mg, 1 mmol,) and heated to 60 deg C. for 2 hours. The
residue was purified by HPLC (Gemini column, 35%
acetonitrile:water, 2 min, 35-50% acetonitrile:water, 2 min,
50-100% acetonitrile: water 13 min, both solvents containing 0.1%
trifluoroacetic acid). This resulted in 37 mg of the title compound
as its TFA salt: MS (m/z): 522.0 [M-H].sup.+; HPLC retention time:
3.58 min (2-98% acetonitrile:water with 0.05% trifluoroacetic
acid).
Example 164
Compound 164--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-((4-trans-methyl-cyclohexanecarbonyl)-{4-tr-
ans-[methyl-(pyridine-3-sulfonyl)-amino]-cyclohexyl}-amino)-thiophene-2-ca-
rboxylic acid
##STR00152##
[0579] (4-trans-Amino-cyclohexyl)-carbamic acid tert-butyl ester
(705 mg, 2.8 mmol) and triethyamine were dissolved in DCM (30 mL)
under an atmosphere of nitrogen and cooled to 0.degree. C. After 10
minutes, pyridine-3-sulfonyl chloride (1.00 g, 5.6 mmol) was added.
The reaction was allowed to warm to room temperature and was
stirred for 18 h. The reaction was found complete by LC/MS. The
reaction was washed with water (2.times.10 mL). The organic layer
was dried with sodium sulfate and filtered.
[4-trans-(pyridine-3-sulfonylamino)-cyclohexyl]-carbamic acid
tert-butyl ester (927 mg, 90%) was purified by precipitation into a
white solid. LC/MS=300 (M.sup.+-55)
[0580] [4-trans-(Pyridine-3-sulfonylamino)-cyclohexyl]-carbamic
acid tert-butyl ester (1.08 g, 3.04 mmol) was dissolved in DMF (30
mL) under an atmosphere of nitrogen. The solution was cooled to
0.degree. C. and Cs.sub.2CO.sub.3 (2.97 g, 9.12 mmol) was added.
After 10 minutes, iodomethane (947 .mu.L, 15.19 mmol) was added.
After 10 minutes, the reaction was allowed to warm to room
temperature. The reaction was stirred for 2.5 h until found
complete by LC/MS. The reaction was quenched with water and
extracted with ethyl acetate (2.times.30 mL). The combined organics
were washed with a 5% LiCl solution in water, dried with sodium
sulfate, filtered and concentrated.
{4-trans-[methyl-(pyridine-3-sulfonyl)-amino]-cyclohexyl}-carbamic
acid tert-butyl ester (741 mg, 66%) was purified by silica gel
chromatography to afford a white solid.
[0581] LC/MS=314 (M.sup.+-55)
[0582]
{4-trans-[Methyl-(pyridine-3-sulfonyl)-amino]-cyclohexyl}-carbamic
acid tert-butyl ester (741 mg, 2.01 mmol) was dissolved in a 4N
solution of HCl in dioxane (2.0 mL, 8.00 mmol). The reaction was
allowed to stir for 30 min until complete by LC/MS. The reaction
was concentrated and used without purification. LC/MS=269
(M.sup.+)
[0583] 5-(3,3-Dimethyl-but-1-ynyl)-3-iodo-thiophene-2-carboxylic
acid methyl ester (469 mg, 1.35 mmol), BINAP (126 mg, 0.2 mmol),
Pd(OAc).sub.2(55 mg, 0.2 mmol) and Cs.sub.2CO.sub.3 (2.20 g, 6.75
mmol) were combined in degassed toluene (15 mL) under an atmosphere
of argon. To the reaction was added the HCl salt of
pyridine-3-sulfonic acid (4-trans-amino-cyclohexyl)-methyl-amide
(crude, 2.01 mmol max). After 15 minutes, the reaction was heated
to 100.degree. C. After 17 h, the reaction was complete by LC/MS.
The reaction was cooled to room temperature and was filtered. The
reaction was concentrated.
5-(3,3-dmethyl-but-1-ynyl)-3-{4-trans-[methyl-(pyridine-3-sulfonyl)-amino-
]-cyclohexylamino}-thiophene-2-carboxylic acid methyl ester (281
mg, 42%) was purified by silica gel chromatography to afford a
yellow solid. LC/MS=490 (M.sup.++1)
[0584]
5-(3,3-Dimethyl-but-1-ynyl)-3-{4-trans-[methyl-(pyridine-3-sulfonyl-
)-amino]-cyclohexylamino}-thiophene-2-carboxylic acid methyl ester
(281 mg, 0.57 mmol) was dissolved in pyridine (5 mL) under an
atmosphere of nitrogen. To the reaction was added
4-trans-methyl-cyclohexanecarbonyl chloride (138 mg, 0.86 mmol).
The reaction was heated to 90.degree. C. for 20 h. The reaction was
cooled to room temperature and was diluted with EtOAc. The reaction
was washed with water (2.times.5 mL). The organic layer was dried
with sodium sulfate, filtered and was concentrated.
5-(3,3-dimethyl-but-1-ynyl)-3-((4-trans-methyl-cyclohexanecarbonyl)-{4-tr-
ans-[methyl-(pyridine-3-sulfonyl)-amino]-cyclohexyl}-amino)-thiophene-2-ca-
rboxylic acid methyl ester (105 mg, 30%) was purified by silica gel
chromatography to afford a yellow solid.
[0585] LC/MS=613 (M.sup.+)
[0586]
5-(3,3-Dimethyl-but-1-ynyl)-3-((4-trans-methyl-cyclohexanecarbonyl)-
-{4-trans-[methyl-(pyridine-3-sulfonyl)-amino]-cyclohexyl}-amino)-thiophen-
e-2-carboxylic acid methyl ester (105 mg, 0.18 mmol) was dissolved
in THF (1 mL) and MeOH (0.5 mL). To the reaction was added a
solution of LiOH (20 mg, 0.88 mmol) in water (1 mL). The reaction
was stirred for 1 h. The reaction was neutralized with 0.88 mL of
1N HCl in water. The solution was concentrated.
5-(3,3-dimethyl-but-1-ynyl)-3-((4-trans-methyl-cyclohexanecarbonyl)-{4-tr-
ans-[methyl-(pyridine-3-sulfonyl)-amino]-cyclohexyl}-amino)-thiophene-2-ca-
rboxylic acid (62 mg, 57) was purified by HPLC to afford a white
powder.
[0587] LC/MS=597 (M.sup.+-2) Neg ionization
[0588] Retention time: 2.62 min
[0589] LC: Thermo Electron Surveyor HPLC
[0590] MS: Finnigan LCQ Advantage MAX Mass Spectrometer
[0591] Column: Phenomenex Polar RP 30 mm.times.4.6 mm
[0592] Solvents: Acetonitrile with 0.1% formic acid, Water with
0.1% formic acid
[0593] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 165
Compound 165--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-[4-(p-
yridine-3-sulfonylamino)-cyclohexyl]-amino}-thiophene-2-carboxylic
acid
##STR00153##
[0595]
3-[(4-Amino-cyclohexyl)-(4-trans-methyl-cyclohexanecarbonyl)-amino]-
-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid methyl
ester (200 mg, 0.44 mmol) and TEA (245 .mu.L, 1.76 mmol) were
dissolved in DCM (3 mL) under an atmosphere of nitrogen and cooled
to 0.degree. C. To the reaction was added pyridine-3-sulfonyl
chloride (155 mg, 0.88 mmol). The reaction was allowed to warm to
room temperature and was stirred for 30 minutes. The reaction was
concentrated.
5-(3,3-dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-[4-(p-
yridine-3-sulfonylamino)-cyclohexyl]-amino}-thiophene-2-carboxylic
acid methyl ester (50 mg, 22%) was purified by silica gel
chromatography to afford a yellow solid. LC/MS=600 (M.sup.++1)
[0596]
5-(3,3-Dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-
-[4-(pyridine-3-sulfonylamino)-cyclohexyl]-amino}-thiophene-2-carboxylic
acid methyl ester (50 mg, 0.1 mmol) was dissolved in a THF (1 mL)
and MeOH (1 mL). To the reaction was added a solution of LiOH (12
mg, 0.5 mmol.) in water (1 mL). The reaction was stirred for 1
hour. The reaction was quenched with 1N HCl in water (0.5 mL).
5-(3,3-Dimethyl-but-1-ynyl)-3-{(4-trans-methyl-cyclohexanecarbonyl)-[4-(p-
yridine-3-sulfonylamino)-cyclohexyl]-amino}-thiophene-2-carboxylic
acid (23 mg, 40%) was purified by HPLC to afford a white
powder.
[0597] LC/MS=584 (M.sup.+-1) Neg ionization
[0598] Retention time: 2.52 min
[0599] LC: Thermo Electron Surveyor HPLC
[0600] MS: Finnigan LCQ Advantage MAX Mass Spectrometer
[0601] Column: Phenomenex Polar RP 30 mm.times.4.6 mm
[0602] Solvents: Acetonitrile with 0.1% formic acid, Water with
0.1% formic acid
[0603] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 186
Compound 186--Synthesis of
3-{4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-methyl-cyc-
lohexanecarbonyl)-amino]-cyclohexylcarbamoyloxy}-azetidine-1-carboxylic
acid tert-butyl ester
##STR00154##
[0605]
3-{4-[[2-Carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-meth-
yl-cyclohexanecarbonyl)-amino]-cyclohexylcarbamoyloxy}-azetidine-1-carboxy-
lic acid tert-butyl ester was prepared in a similar fashion to
4-[[2-carboxy-5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-trans-methyl--
cyclohexanecarbonyl)-amino]-piperidine-1-carboxylic acid
piperidin-4-yl ester using method A except that the HCl salt of
3-[(4-amino-cyclohexyl)-(4-trans-methyl-cyclohexanecarbonyl)-amino]-5-(3,-
3-dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid was used instead
of the HCl salt of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-trans-methyl-cyclohexanecarbonyl)-piper-
idin-4-yl-amino]-thiophene-2-carboxylic acid and
3-hydroxy-azetidine-1-carboxylic acid tert-butyl ester was used
instead of 4-hydroxy-piperidine-1-carboxylic acid tert-butyl
ester.
[0606] LC/MS (m/z): 544 [M-99]
[0607] Retention time: 2.52 min
[0608] LC: Thermo Electron Surveyor HPLC
[0609] MS: Finnigan LCQ Advantage MAX Mass Spectrometer
[0610] Column: Phenomenex Polar RP 30 mm.times.4.6 mm
[0611] Solvents: Acetonitrile with 0.1% formic acid, Water with
0.1% formic acid
[0612] Gradient: 0 min-0.1 min 5% ACN, 0.1 min-1.95 min 5%-100%
ACN, 1.95 min-3.5 min 100% ACN, 3.5 min-3.55 min 100%-5% ACN, 3.55
min-4 min 5% ACN.
Example 217
Compound 217--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-((4-methyl-cyclohexanecarbonyl)-{4-[N'-meth-
yl-N'-(tetrahydro-pyran-4-yloxycarbonyl)-hydrazino]-cyclohexyl}-amino)-thi-
ophene-2-carboxylic acid
##STR00155##
[0614] To a solution of tetrahydro-pyran-4-ol (0.215 mL, 2.27 mmol)
and N,N'-disuccimidyl carbonate (0.87 g, 3.4 mmol) in ACN (7 mL)
was added triethylamine (1 mL, 6.81 mmol). After 16 h stirring at
rt, the reaction was quenched with LiCl solution (5% in water, 10
mL) and diluted with EtOAc (20 mL). The crude product was washed
2.times.10 mL with 5% LiCl solution, dried over Na.sub.2SO.sub.4
and concentrated. The crude material was used as is for the next
step.
[0615] A mixture of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-methyl-cyclohexanecarbonyl)-(4-oxo-cycl-
ohexyl)-amino]-thiophene-2-carboxylic acid (100 mg, 0.23 mmol),
N-methyl-hydrazinecarboxylic acid tetrahydro-pyran-4-yl ester (70
mg, .about.0.4 mmol), acetic acid (3 drops) in DCE (3 mL) was
treated with NaBH(OAc).sub.3 (12 mg, 56 mmol) for 16 h. The
reaction was quenched with water (10 mL) and extracted with EtOAc.
The crude material was then purified by HPLC (Gemini column, 35%
acetonitrile:water, 2 min, 35-50% acetonitrile:water, 2 min,
50-100% acetonitrile: water 13 min, both solvents containing 0.1%
trifluoroacetic acid) giving the title compound as a TFA salt: MS
(m/z): 603.1 [M-H]'; HPLC retention time: 27.4 min (2-98%
acetonitrile:water with 0.05% trifluoroacetic acid) 30 min run.
Example 218
Compound 218--Synthesis of
5-(3,3-dimethyl-but-1-ynyl)-3-((4-methyl-cyclohexanecarbonyl)-{4-[N'-meth-
yl-N'-(pyridin-2-yloxycarbonyl)-hydrazino]-cyclohexyl}-amino)-thiophene-2--
carboxylic acid
##STR00156##
[0617] To a solution of carbonic acid di-2-pyridyl ester (300 mg,
1.38 mmol) in MeOH (10 mL) was added methylhydrazine (94 uL, 1.8
mmol). After 16 h stirring at rt, the reaction was concentrated and
the crude material was used as is for the next step.
[0618] A mixture of
5-(3,3-dimethyl-but-1-ynyl)-3-[(4-methyl-cyclohexanecarbonyl)-(4-oxo-cycl-
ohexyl)-amino]-thiophene-2-carboxylic acid (100 mg, 0.23 mmol),
N-methyl-hydrazinecarboxylic acid pyridin-2-yl ester (67 mg,
.about.0.4 mmol), acetic acid (3 drops) in DCE (3 mL) was treated
with NaBH(OAc).sub.3 (12 mg, 56 mmol) for 16 h. The reaction was
quenched with water (10 mL) and extracted with EtOAc. The crude
material was then purified by HPLC (Gemini column, 35%
acetonitrile:water, 2 min, 35-50% acetonitrile:water, 2 min,
50-100% acetonitrile: water 13 min, both solvents containing 0.1%
trifluoroacetic acid) giving the title compound as a TFA salt: MS
(m/z): 595.1 [M-H].sup.+; HPLC retention time: 28.9 min (2-98%
acetonitrile:water with 0.05% trifluoroacetic acid) 30 min run.
Biological Examples
Antiviral Activity
[0619] Another aspect of the invention relates to methods of
inhibiting viral infections, comprising the step of treating a
sample or subject suspected of needing such inhibition with a
composition of the invention.
[0620] Within the context of the invention samples suspected of
containing a virus include natural or man-made materials such as
living organisms; tissue or cell cultures; biological samples such
as biological material samples (blood, serum, urine, cerebrospinal
fluid, tears, sputum, saliva, tissue samples, and the like);
laboratory samples; food, water, or air samples; bioproduct samples
such as extracts of cells, particularly recombinant cells
synthesizing a desired glycoprotein; and the like. Typically the
sample will be suspected of containing an organism which induces a
viral infection, frequently a pathogenic organism such as a tumor
virus. Samples can be contained in any medium including water and
organic solvent\water mixtures. Samples include living organisms
such as humans, and man made materials such as cell cultures.
[0621] If desired, the anti-virus activity of a compound of the
invention after application of the composition can be observed by
any method including direct and indirect methods of detecting such
activity. Quantitative, qualitative, and semiquantitative methods
of determining such activity are all contemplated. Typically one of
the screening methods described above are applied, however, any
other method such as observation of the physiological properties of
a living organism are also applicable.
[0622] The antiviral activity of a compound of the invention can be
measured using standard screening protocols that are known. For
example, the antiviral activity of a compound can be measured using
the following general protocols.
Cell-Based Flavivirus Immunodetection Assay
[0623] BHK21 or A549 cells are trypsinized, counted and diluted to
2.times.10.sup.5 cells/mL in Hams F-12 media (A549 cells) or
RPMI-1640 media (BHK21 cells) supplemented with 2% fetal bovine
serum (FBS) and 1% penicillin/streptomycin. 2.times.10.sup.4 cells
are dispensed in a clear 96-well tissue culture plates per well and
placed at 37.degree. C., 5% CO.sub.2 overnight. On the next day,
the cells are infected with viruses at multiplicity of infection
(MOI) of 0.3 in the presence of varied concentrations of test
compounds for 1 hour at 37.degree. C. and 5% CO.sub.2 for another
48 hours. The cells are washed once with PBS and fixed with cold
methanol for 10 min. After washing twice with PBS, the fixed cells
are blocked with PBS containing 1% FBS and 0.05% Tween-20 for 1
hour at room temperature. The primary antibody solution (4G2) is
then added at a concentration of 1:20 to 1:100 in PBS containing 1%
FBS and 0.05% Tween-20 for 3 hours. The cells are then washed three
times with PBS followed by one hour incubation with horseradish
peroxidase(HRP)-conjugated anti-mouse IgG (Sigma, 1:2000 dilution).
After washing three times with PBS, 50 microliters of
3,3',5,5'-tetramethylbenzidine (TMB) substrate solution (Sigma) is
added to each well for two minutes. The reaction is stopped by
addition of 0.5 M sulfuric acid. The plates are read at 450 nm
abosorbance for viral load quantification. After measurement, the
cells are washed three times with PBS followed by incubation with
propidium iodide for 5 min. The plate is read in a Tecan Safire.TM.
reader (excitation 537 nm, emission 617 nm) for cell number
quantification. Dose response curves are plotted from the mean
absorbance versus the log of the concentration of test compounds.
The EC.sub.50 is calculated by non-linear regression analysis. A
positive control such as N-nonyl-deoxynojirimycin may be used.
Cell-Based Flavivirus Cytopathic Effect Assay
[0624] For testing against West Nile virus or Japanese encephalitis
virus, BHK21 cells are trypsinized and diluted to a concentration
of 4.times.10.sup.5 cells/mL in RPMI-1640 media supplemented with
2% FBS and 1% penicillin/streptomycin. For testing against dengue
virus, Huh7 cells are trypsinized and diluted to a concentration of
4.times.10.sup.5 cells/mL in DMEM media supplemented with 5% FBS
and 1% penicillin/streptomycin. A 50 microliter of cell suspension
(2.times.10.sup.4 cells) is dispensed per well in a 96-well optical
bottom PIT polymer-based plates (Nunc). Cells are grown overnight
in culture medium at 37.degree. C., 5% CO.sub.2, and then infected
with West Nile virus (e.g. B956 strain) or Japanese encephalitis
virus (e.g. Nakayama strain) at MOI=0.3, or with dengue virus (e.g.
DEN-2 NGC strain) at MOI=1, in the presence of different
concentrations of test compounds. The plates containing the virus
and the compounds are further incubated at 37.degree. C., 5%
CO.sub.2 for 72 hours. At the end of incubation, 100 microliters of
CellTiter-Glo.TM. reagent is added into each well. Contents are
mixed for 2 minutes on an orbital shaker to induce cell lysis. The
plates are incubated at room temperature for 10 minutes to
stabilize luminescent signal. Lumnescence reading is recorded using
a plate reader. A positive control such as N-nonyl-deoxynojirimycin
may be used.
Antiviral Activity in a Mouse Model of Dengue Infection.
[0625] Compounds are tested in vivo in a mouse model of dengue
virus infection (Schul et al. J. Infectious Dis. 2007; 195:665-74).
Six to ten week old AG129 mice (B&K Universal Ltd, Hll, UK) are
housed in individually ventilated cages. Mice are injected
intraperitoneally with 0.4 mL TSV01 dengue virus 2 suspension.
Blood samples are taken by retro orbital puncture under isoflurane
anaesthesia. Blood samples are collected in tubes containing sodium
citrate to a final concentration of 0.4%, and immediately
centrifuged for 3 minutes at 6000 g to obtain plasma. Plasma (20
microliters) is diluted in 780 microliters RPMI-1640 medium and
snap frozen in liquid nitrogen for plaque assay analysis. The
remaining plasma is reserved for cytokine and NS1 protein level
determination. Mice develop dengue viremia rising over several
days, peaking on day 3 post-infection.
[0626] For testing of antiviral activity, a compound of the
invention is dissolved in vehicle fluid, e.g. 10% ethanol, 30% PEG
300 and 60% D5W (5% dextrose in water; or 6N HCl (1.5 eq):1N NaOH
(pH adjusted to 3.5): 100 mM citrate buffer pH 3.5 (0.9% v/v:2.5%
v/v: 96.6% v/v). Thirty six 6-10 week old AG129 mice are divided
into six groups of six mice each. All mice are infected with dengue
virus as described above (day 0). Group 1 is dosed by oral gavage
of 200 mL/mouse with 0.2 mg/kg of a compound of the invention twice
a day (once early in the morning and once late in the afternoon)
for three consecutive days starting on day 0 (first dose just
before dengue infection). Groups 2, 3 and 4 are dosed the same way
with 1 mg/kg, 5 mg/kg and 25 mg/kg of the compound, respectively. A
positive control may be used, such as
(2R,3R,4R,5R)-2-(2-amino-6-hydroxy-purin-9-yl)-5-hydroxymethyl-3-methyl-t-
etrahydro-furan-3,4-diol, dosed by oral gavage of 200
microliters/mouse the same way as the previous groups. A further
group is treated with only vehicle fluid.
[0627] On day 3 post-infection approximately 100 microliter blood
samples (anti-coagulated with sodium citrate) are taken from the
mice by retro-orbital puncture under isoflurane anaesthesia. Plasma
is obtained from each blood sample by centrifugation and snap
frozen in liquid nitrogen for plague assay analysis. The collected
plasma samples are analyzed by plague assay as described in Schul
et al. Cytokines are also analysed as as described by Schul. NS1
protein levels are analysed using a Platelia.TM. kit (BioRad
Laboratories). An anti-viral effect is indicated by a reduction in
cytokine levels and/or NS1 protein levels.
[0628] Typically, reductions in viremia of about 5-100 fold, more
typically 10-60 fold, most typically 20-30 fold, are obtained with
5-50 mg/kg bid dosages of the compounds of the invention.
HCV Assay Protocol
[0629] The anti-HCV activity of the compounds of this invention was
tested in a human hepatoma Huh-7 cell line harboring a HCV
replicon. The assay comprised the following steps:
Step 1: Compound Preparation and Serial Dilution.
[0630] Serial dilution was performed in 100% DMSO in a 384-well
plate. A solution containing a compound at 225-fold concentration
of the starting final serial dilution concentration was prepared in
100% DMSO and 15 uL added to the pre-specified wells in column 3 or
13 of a polypropylene 384-well plate. The rest of the 384-well
plate was filled with 10 uL 100% DMSO except for columns 23 and 24,
where 10 uL of 500 uM a HCV protease inhibitor (ITMN-191) in 100%
DMSO was added. The HCV protease inhibitor was used a control of
100% inhibition of HCV replication. The plate was then placed on a
Biomek FX Workstation to start the serial dilution. The serial
dilution was performed for ten cycles of 3-fold dilution from
column 3 to 12 or from column 13 to 22.
Step 2: Cell Culture Plate Preparation and Compound Addition
[0631] To each well of a black polypropylene 384-well plate, 90
.mu.L of cell media containing 1600 suspended Huh-7 HCV replicon
cells was added with a Biotek uFlow Workstation. A volume of 0.4
.mu.L of the compound solution was transferred from the serial
dilution plate to the cell culture plate on a Biomek FX
Workstation. The DMSO concentration in the final assay condition
was 0.44%. The plates were incubated for 3 days at 37.degree. C.
with 5% CO2 and 85% humidity.
Step 3: Detection of Cytotoxicity and Inhibition of Viral
Replication
[0632] a) Assessment of cytotoxicity: The media in the 384-well
cell culture plate was aspirated with a Biotek EL405 plate-washer.
A volume of 50 .mu.L of a solution containing 400 nM Calcein AM in
100% PBS was added to each well of the plate with a Biotek uFlow
Workstation. The plate was incubated for 30 minutes at room
temperature before the fluorescence signal (emission 490 nm,
exitation 520 nm) was measured with a Perkin Elmer Envision Plate
Reader.
[0633] b) Assessment of inhibition of viral replication: The
calcein-PBS solution in the 384-well cell culture plate was
aspirated with a Biotek EL405 plate-washer. A volume of 20 .mu.L of
Dual-Glo luciferase buffer (Promega, Dual-Glo Luciferase Assay
Reagent, cat. #E298B) was added to each well of the plate with a
Biotek uFlow Workstation. The plate was incubated for 10 minutes at
room temperature. A volume of 20 .mu.L of a solution containing
1:100 mixture of Dual-Glo Stop & Glo substrate(Promega,
Dual-Glo Luciferase Assay Reagent, cat. #E313B) and Dual-Glo Stop
& Glo buffer (Promega, Dual-Glo Luciferase Assay Reagent, cat.
#E314B) was then added to each well of the plate with a Biotek
uFlow Workstation. The plate was incubated at room temperature for
10 minutes before the luminescence signal was measured with a
Perkin Elmer Envision Plate Reader.
Step 4: Calculation
[0634] The percent cytotoxicity was determined by calcein AM
conversion to fluorescent product. The average fluorescent signal
from the DMSO control wells were defined as 100% nontoxic. The
individual fluorescent signal from testing compound treated well
was divided by the average signal from DMSO control wells and then
multiplied by 100% to get the percent viability. The percent
anti-HCV replication activity was determined by the luminescence
signal from the testing well compared to DMSO controls wells. The
background signal was determined by the average luminescence signal
from the HCV protease inhibitor treated wells and was subtracted
from the signal from the testing wells as well as the DMSO control
wells. Following 3-fold serial dilutions, the EC.sub.50 and
CC.sub.50 values were calculated by fitting % inhibition at each
concentration to the following equation:
% inhibition=100%[(EC.sub.50/[l]).sup.b+1]
[0635] Where b is Hill's coefficient. See, for reference, Hill, A.
V., The Possible Effects of the Aggregation of the Molecules of
Hmoglobin on its Dissociation Curves, J. Physiol. 40: iv-vii.
(1910).
[0636] % inhibition values at a specific concentration, for example
2 .mu.M, can also be derived from the formula above.
[0637] When tested, certain compounds of this invention were found
to inhibit viral replication as listed in Table 1:
TABLE-US-00002 TABLE 1 Compound % inhibition at 2 .mu.M 31 93.1 32
100 33 96.2 34 97.8 35 99.8 36 100 37 99.6 38 99.6 39 98.8 40 98.6
41 99.9 42 99.9 43 96.2 44 100 45 100 46 97.0 47 99.9 48 100 49
99.2 50 99.9 51 85.5 52 93.5 53 99.9 54 99.8 55 87.9 56 92.8 57
78.2 64 100 65 99.9 66 99.9 67 99.0 68 98.5 69 99.5 70 99.1 71 95.1
72 99.7 73 93.1 74 99.7 75 97.5 76 99.4 77 99.9 79 100 80 99.8 81
100 82 100 83 99.9 112 99.9 113 98.1 114 100 115 99.8 116 99.9 117
99.9 118 100 119 97.1 120 99.9 121 99.9 122 99.7 123 99.9 124 99.9
125 99.7 126 98.9 127 98.3 128 99.8 129 100 130 98.7 131 99.9 132
77.4 133 96.7 134 99.0 135 99.8 136 98.9 137 100 140 99.9 141 99.9
142 99.6 164 98.9 165 95.7 186 98.9 217 100 218 100
[0638] Preferred compounds according to Table 1 include Examples
32, 36, 38, 41, 48, 50, 64, 65, 66, 77, 79, 80, 81, 82, 83, 112,
114, 118, 129, 217, and 218.
[0639] The specific pharmacological responses observed may vary
according to and depending on the particular active compound
selected or whether there are present pharmaceutical carriers, as
well as the type of formulation and mode of administration
employed, and such expected variations or differences in the
results are contemplated in accordance with practice of the present
invention.
[0640] Although specific embodiments of the present invention are
herein illustrated and described in detail, the invention is not
limited thereto. The above detailed descriptions are provided as
exemplary of the present invention and should not be construed as
constituting any limitation of the invention. Modifications will be
obvious to those skilled in the art, and all modifications that do
not depart from the spirit of the invention are intended to be
included with the scope of the appended claims.
* * * * *